AU681875B2 - 2-benzyl-polycyclic guanine derivatives and process for preparing them - Google Patents

Description

OPI DATE 14/09/94 APPLN. ID 62687/94 l!llllII!II ll IIHI 11 1f illi r' AOJP DATE 17/11/94 PCT NtJNI3R PCT/US94/01728 1111111111 N11 11l111,1 11111 El1111UI1I li111 IEI1I1III 111l AtI9462637

DITL

WO 94119351 (51) International Patent Classification 5 C071) 487/14, A61K( 311505, C07D 487/20, 491/22, 473118, 473/40, 239/54, 239/52 fl (CO7D 487/14, 239:00, 235:00, 235:00) (CO7D 49-/20, 239:00, 235:00, 235:00) (COW 491122, 307:00, 239:00, 235:00, 235:00) (11) International Publication Number: (43) International Publication Date: 1 September 1994 (01.09.94) (21) International Application Number: (22) International Filing Date: 24 Priority Data.

08/024,599 26 February 08/023,549 26 February Parent Applications or Grants (63) Related by Continuation us Filed on 26 us Filed on 26: PCTIUS94/01728 February 1994 (24.02.94) *1993 (26.02.93) 1993 (26.02,93) 08/023,549 (ClIP) February 1993 (26.02.93) 08/024,599 (CII') February 1993 (26.02.93) (72) Inventors; and (75) Inventors/Applicants (for US only): TULSUN, Dean rN/US]; 46 Oakland Avenue, Rockaway, NJ 07866 (US).

MoK1lTRCK, Brian, A- [US/US]; 67 Laurel Avenue, Bloomfield, NJ 07003 XLA, Yan [CN/USI; 137 Christie Street, Edison, NJ 08820 CHACKALAMAN- NIL. Samuel [IN/US]; 79 Stratford Road, East Brun~swick, NJ 08816 GALA, Dinesh CUS/US];- 9 Sylvester Court East Brunswick4 NJ 08816 DIBENEDETIO, Donald, J. [US/US]; 209 Pcmbrokl Road, Mountainside, NJ 07092 KUGELMAN, Max [US/US]; 271 Elsinre Place, Fort Lee, NJ 07024 (US).

(74) Agents: MAGATTI Anita, W. et al.; Schering-~Plough Corporation, One Giralda Farms, ?MW, Madison, NJ 07940-1000

(US).

(81) Designated States: AU, BB, BG, BR, BY, CA, CN, CZ, Fl, HU, 1P, KR, KZ, LV, MG, AN, MW, NO, NZ, PL, RO, RU, SD, UA, US, UZ, VN, European patent (AT, BE, CHI, DE, ES, FR, GB, GR, IE, rT, LU, MC, Nb, PT, SE), GAPI patent (BF, BJ, CF, CG, CL, CM GA, GN, ML, MR NE. SN, TD, TG).

Published With international search report.

Before the expiration of the time limit for amending the claims and to be republished in the event of the receipt of amendment.

(71) Applicant (for all designated States except US): SCHERING CORPORATION [US/US]; 2000 Galloping Hill Road, Keailworth, NJ 07033 (US).

(54) Title: 2-BENZYL-POLYCYCLIC GUANINE DERIVATIVES AND PROCESS FOR PREPARING THEM (57) Abstract Antihypertensive and0 bronchodilating compounds of H formula or a pharmaceutically N

R

aprocess for preparing them CHXR are disclosed, wherein: Ri, R2 and R 3 are hydrogen, lower NR alkyl, lower alkoxty, halogeno, hydroxy, (di-lower alkyl)ammno, f' R 4..morpholinyl, 1-pyrrolidinyl, R b 1-pyrrolyL, -CF 3 .0CF 3 phenyl or methoxyphenyl; or Rt and R-2 together are methylenedioxy; or R1 and R2 together with the carbon atoms to which they are attached form a benzene ring-, and R4 is hydrogen and Rb and together with the carbon atoms to which they are attached, form a saturated ring of 5 carbons; or Ra is lower alkyl, Rb is hydrogen or lower alkyl, and RO is hydrogen; or R3, R11 and the carbxn atom to which they are attached form a saturated ring of 5-7 carbons, and Rc is hydrogen; or Ra is hydrogen, and Rb, R 0 and the carbon atoms to which they are attached form a tetrahydrofuran ring, or RI and Rb, together with the carbon atom to which they are attached, and Rb and Re, together with the carbon atoms to wbich they are attached, each form a saturated ring of 5-7 carbons; pharmaceutical compositions containing said compounds, methods of treatment using said icompounds; and a process for preparing polycydlic guanines comprising a) reducing a nitrosopyrimidine, and treating the reduced nitrosopyrimnidine with an acylating reagent to give the aniidopyrimidine; b) reacting the axnidopyi-imidine with a halogenating/cyclizing reagent to give a halopurine; c) reacting, in the presence of a base, the halopurine with an amine to give the substituted a'iopurine; and A) closin the ring of the bstituted aminopurine with a suitAble dehydraing agent _I I SWO 94/19351 PCT/US94101728 2-BENZYL-POLYCYC LIC GUANINE DERIVATIVES AND PROCESS FOR PREPARING THEM

BACKGROUND

The present invention relates to 2-benzyl-polycyclic guanine derivatives useful for treating cardiovascular and pulmonary disorders, as well as to their pharmaceutical compositions, methods for using the same and a process for preparing them. Most compounds of this invention were generically but not specifically disclosed in PCT publication W091/19717, published December 26, 1991. We have found that the compounds of the present invention show unexpectedly superior cardiovascular and pulmonary activity compared to the compounds of the prior publication.

SUMMARY OF THE INVENTION The present invention is directed to novel 2-benzyl-polycyclic guanine derivatives of the formula: 0

H

C N -d-R 2 a N R3 Rb I or a pharmaceutically acceptable salt thereof, whereii:

R

1

R

2 and R 3 are independently selected from the group consisting of hydrogen, lower alkyl, lower alkoxy, halogeno, hydroxy, (dilower alkyl)amino, 4-morpholinyl, 1-pyrrolidinyl, 1-pyrrolyl, -CF 3 -OCF3, phenyl and methoxyphenyl; or Ri and R 2 together are methylenedioxy; or R 1 and R 2 together with the carbon atoms to which they are attached form a benzene ring; and

R

a is hydrogen and Rb and Rc, together with the carbon atoms to which they are attached, form a saturated ring of 5 carbons; or R a U IL 1 ~1~4s I_ I SWO 94/19351 PCT/US94/01728 -2is lower alkyl, Rb is hydrogen or lower alkyl, and RC is hydrogen; or R a Rb and the carbon atom to which they are attached form a saturated ring of 7 carbons, and Rc is hydrogen; or R a is hydrogen, and Rb, Rc and the carbon atoms to which they are attached form a tetrahydrofuran ring; or R a and Rb, together with the carbon atom to which they are attached, and Rb and Rc, together with the carbon atoms to which they are attached, each form a saturated ring of 5-7 carbons.

Preferred compounds are those wherein R1 and R 3 are hydrogen. More preferred are compounds wherein R1 and R 3 are hydrogen and R 2 is hydrogen, -OCF 3 methyl, methoxy, fluoro, phenyl, methoxyphenyl, dimethylamino, 1-pyrrolidinyl or 1-pyrrolyl, and compounds wherein R 1 and R 2 together are methylenedioxy and R 3 is hydrogen.

Also preferred are compounds wherein R a is hydrogen and Rb and Rc, together with the carbon atoms to which they are attached, form a saturated ring of 5 carbons; compounds wherein R a is lower alkyl, Rb is hydrogen or lower alkyl, and R c is hydrogen; compounds wherein R a and Rb and the carbon atom to which they are attached form a saturated ring of 5 carbons and RC is hydrogen; compounds wherein R a is hydrogen and Rb, RC and the carbon atoms to which they are attached form a tetrahydrofuran ring; and compounds wherein R a and Rb, together with the carbon atoms to which they are attached, and Rb and RC, together with the carbon atoms to which they are attached, each form a saturated ring of 5 carbons. More preferred are compounds wherein R a is hydrogen and Rb and Rc, together with the carbon atoms to which they are attached, form a saturated ring of 5 carbons; compounds wherein R a and Rb and the carbon atom to which they are attached form a saturated ring of 5 carbons and RC is hydrogen; and compounds wherein R a is lower alkyl, Rb is hydrogen or lower alkyl, and Rc is hydrogen.

Of compounds of formula I wherein R a is hydrogen and R b and Rc, together with the carbon atoms to which they are attached, form a saturated ring of 5 carbons, most preferred are compounds wherein R 1 R2 and R 3 are as listed in the following table: a aab~ Ibs I WO 94/19351 PCT[UJS94/01728 -3- Ri R 2 R3- H H H H F H H -OCH 3

H

H -CHs H H (CH 3 2 N- H H C 6

H

s

H

H -OCF 3

H

H -N H H N H 3,4-OCH 2 0- H H -C 6

H

4 0CH3 H The compounds of formula I are useful as antihypertensive, bronchodilating and blood platelet inhibiting agents. Compounds of the invention are useful in inhibiting phosphodiesterase enzymes; the inhibition of vascular phosphodiesterase is associated with vasodilation and vasorelaxation, and therefore is expected to induce antihypertensive and antianginal activity. Compounds of formula I can also serve as smooth muscle relaxants and are therefore useful in the treatment of bronchoconstriction. Such compounds also can inhibit smooth muscle proliferation, vascular growth and platelet function and are useful in treating conditions such as restenosis post angioplasty, atherosclerosis and conditions wnich benefit from inhibiting platelet function. Through one or more of the above physiological mechanisms, compounds of formula I are also useful in treating ischemia and peripheral vascular diseases.

The present invention is also directed toward a pharmaceutical composition containing a compound of formula I in an amount effective to inhibit phosphodiesterase enzymes, smooth muscle proliferation, vascular growth or platelet function, or to relax smooth muscle. The present invention is also directed toward a pharmaceutical composition containing an anti-hypertensive, an anti-anginal, a bronchodilating or a platelet inhibiting effective amount of a compound of formula I.

The present invention is also directed toward a method for treating hypertension, angina, bronchoconstriction, restenosis post 9 1 ~b C I I _II- WO 94/19351 PCT/US94/01728 -4angioplasty, atherosclerosis, ischemia, peripheral vascular diseases, or diseases benefitting from platelet inhibition in a mammal comprising administering to a mammal in need of such treatment an amount of a compound of formula I iffective to treat any of the above diseases. The present invention is also directed toward a method for maintaining guanosine 3':5'-cyclic monophosphate (cGMP) levels in a mammal by administering an amount of a compound of formula I effective to maintain or increase cGMP levels.

In another embodiment, the present invention is directed toward the preparation of a polycyclic guanine of formula I o R4

H

N N RN

N

Ral (CH)n Rbl RC1 Rdi wherein,

R

4 is H, alkyl or alkyl substituted with aryl or -OH;

R

5 is H, halo, -CF 3 alkoxy, alkylthio, alkyl, cycloalkyl, -SO 2

NH

2

-NH

2 monoalkylamino, dialkylamino, hydroxyalkylamino, aminoalkylamino, -COOH, alkoxycarbonyl, aminocarbonyl, aryl, substituted aryl or alkyl substituted with aryl, substituted aryl, -OH, alkoxy, -NH 2 monoalkylamino or dialkylamino; Ral, R b l, R cl and Rdl independently represent H, alkyl, cycloalky! or aryl; or (R a l and Rbl) or (R cl and Rdl) or (Rbl and R 0 1 can complete a saturated ring of 5- to 7- carbon atoms, or (R al and R b l) taken together and (Rbl and R cl taken together, each complete a saturated ring of 5- to 7-carbon atoms, wherein each ring optionally can contain a sulfur or oxygen atom and whose carbon atoms may be optionally substituted with one or more or the following: alkenyl, alkynyl, -OH, -COOH, alkoxycarbonyl, alkyl or alkyl substituted with -OH, -COOH or alkoxycarbonyl; or such saturated ring can have two adjacent carbon atoms which are shared with an adjoining aryl ring; and n is zero or one; wherein the process comprises: a) reducing a nitrosopyrimidine (III), and treating the reduced nitrosopyrimidine with an acylating reagent optionally in the L L~s C ~a~L ~a~~LIC~t4- 111 IWO 94/19351 PCT/US94/01728 presence of either an acylating catalyst and/or coupling reagent and/or phase transfer catalyst to give the amidopyrimidine b) reacting the amidopyrimidine with an effective amount of a halogenating/cyclizing reagent, optionally in the presence of one or more additional halide sources, and also optionally in the presence of a phase transfer catalyst to give ai halopurine (VI); c) reacting, in the presence of a base, the halopurine (VI) with an amine of the formula:

HN

a

(CH

2 n

-OH

Rbl Rl Rdl (VII) wherein Ral Rbl, R

C

Rdl and n are as described above to give the substituted aminopurine (VIII); d) closing the ring of the substituted aminopurine (VIII) with a suitable dehydrating agent to give the polycyclic guanine (II).

In another embodiment, the present invention is directed toward a process for preparing amidopyrimidines of formula V, comprising: reducing a nitrosopyrimidine of formula III, and treating the reduced nitrosopyrimidine with an acylating reagent, optionally in the presence of either an acylating catalyst and/or coupling reagent and/or phase transfer catalyst. This process corresponds to step a) above.

In another embodiment, the present invention is directed toward a process for preparing a halopurine of formula VI, comprising reacting an amidopyrimidine with an effective amount of a halogenating/cyclizing reagent, optionally in the presence of one or more additional halide sources, and also optionally in the presence of a phase transfer catalyst. This process corresponds to step above. This process can further include step and/or step described below.

In another embodiment, the present invention is directed toward a process for preparing a substituted aminopurine of formula VIII, comprising reacting, in the presence of a base, the halopurine (VI) with an amine of the formula:

H

2

N

S\ (CH2)n-OH Ral Rb 1 Re R VI ~s -1 I r g~p~pg -IY1 I I WO 94/19351 PCTUS94/01728 -6wherein R a l Rbl, R c l Rdl and n are as described above. This process corresponds to step c) above. This process can further comprise step described below.

The present invention is also directed toward the novel intermediates: amidopyrimidine V, halopurine VI and aminopurine VIII, with the proviso that with regards to amidopyrimidine V, where Z is R 4 and R 5 cannot both be methyl.

The present invention has the advantage of providing a -ocess for preparing polycylic guanine derivatives and intermediates thereof, in as few or even fewer steps than other processes previously taught, in good yields with little formation of undesirable by-products, with less waste to dispose of or recycle. The present invention has the further advantage of providing novel intermediates which enable the above process to achieve these advantages.

DETAILED DESCRIPTION OF THE INVENTION In describing the present invention, "lower alkyl" represents a straight alkyl chain having from 1 to 6 carbon atoms or a branched alkyl chain of 3 to 6 carbon atoms, for example methyl, ethyl, propyl, isopropyl, butyl, isobutyl, tert-butyl, pentyl and hexyl.

"Lower alkoxy" represents an alkoxy group wherein the alkyl portion is as defined above, for example, methoxy, ethoxy, propoxy, pentyloxy and hexyloxy.

"Halogeno" represents fluoro, chloro, bromo or iodo.

"Aryl" represents a carbocyclic moiety containing at least one benzenoid-type ring, with the aryl moiety having from 6 to 14 carbon atoms, with all available substitutable carbon atoms of the aryl moiety being intended as possible points of attachment, for example phenyl, naphthyl, indenyl, indanyl and the like.

"Substituted aryl" represents an aryl moiety as defined above substituted with 1 to 3 substituents selected from the group consisting of halogeno, lower alkyl, -CF 3

-OCF

3 phenyl, -OH, lower alkoxy, phenoxy, amino, (mono-lower alkyl)amino, (di-lower alkyl)amino, 4-morpholinyl, 1pyrrolidinyl, 1-pyrrolyl and methoxyphenyl, or substituents on adjacent carbon atoms form a methylenedioxy group.

Certain compounds of the invention those with a basic nitrogen containing moiety, can also form pharmaceutically acceptable salts with organic and inorganic acids. Examples of suitable acids for ~aasl IPI r~~l~ I I -I u WO 94/19351 PCT/US94/01728 -7such salt formation are hydrochloric, sulfuric, phosphoric, acetic, citric, oxalic, malonic, salicylic, malic, fumaric, succinic, ascorbic, maleic, methanesulfonic and other mineral and carboxylic acids w :own to those skilled in the art. The salts are prepared by contact'- ;e free base form with a sufficient amount of the desired acid to produ., a salt in the conventional manner.

Certain compounds of the invention will be acidic in nature, those compounds which possess a carboxy or phenolic hydroxyl group. These compounds may form pharmaceutically acceptable salts.

Examples of such salts are the sodium, potassium, calcium, aluminum, gold and silver salts. Also contemplated are salts formed with pharmaceutically acceptable amines such as ammonia, alkylamines, hydroxyalkylamines, N-methylglucamine and the like.

Compounds of formula I form enantiomers, with the enantiomeric form being preferred. For example, the preferred stereochemistry for compounds wherein R a is hydrogen and Rb and RC, together with the carbon atoms to which they are attached, form a saturated ring of 5 carbons is shown in the following partial structural formula: N, N The process aspect of the present invention and its various embodiments are illustrated below: O O R4. Reduction/ R4 R4 NO RR0OL IV "N 4

NHCOR

s N3 o RsCOL IV N .2 6 !6L r11s Z N NH 2 Z N NH 2 III

V

0 4 H 0 N N R 4 .N H N R s VII

NR

s HN<N

N

S (CH-)n-OH X N R Rol Rdi VIII ~a ne recJ~i~ r~a~ WO 94/19351 PCT/US94/01728 -8- 0 R4 J H N N VIII N Rai (CH2)n Rh RC R II wherein R 4

R

5 Ral Rbl, Rd l Rdl and n are as defined above; L is a leaving group; X is halogeno; Z is =0 or -OR 6 wherein R 6 is alkyl; and the dotted lines indicate an optional double bond such that when Z is position 1 contains hydrogen and there is single bond 9etween positions 1 and 2 on the ring, or when Z is -OR 6 wherein R 6 is alkyl, there is a double bond between positions 1 and 2 on the ring; The amidopyrimidines of formula V can be prepared by reducing a nitrosopyrimidines of formula III, followed by treatment with an acylating reagent. Suitable reducing agents include hydrogen with a metal catalyst, such as the metals from Group VIII of the periodic table or salts or complexes thereof, or a mixture of metal of Group VIII with carbon.

Suitable metals include platinum, palladium, nickel, rhodium, ruthenium or mixtures thereof. The reduced nitrosopyrimidine is then treated with an acylating reagent, optionally in the presence of either an acylating catalyst and/or coupling reagent and/or phase transfer catalyst. In the reduction of nitrosopyrimidines, the metal catalyst can be employed in amounts effective to give the reduced nitrosopyrimidine. Such amounts can range from about 1 to about 200 mole percent of the metal catalyst, preferably from about 1 to about 50 mole percent, more preferably about 1 to about mole percent metal catalyst. Reduction can be carried out at temperatures effective to give th' the reduced nitrosopyrimidine, and can range from about -400C to abo' 100°C, more preferably from about 00 to about 50°C, more preferably r. mn 100 to 300C. The nitrosopyrimidine of formula III is reduced under pressures ranging from about ambient to about 400 pounds per square inch (psi) (20 kilotorr), preferably from about 35 psi (1.8 kilotorr) to 100 psi (5.1 kilotorr), more preferably from about 50 psi (2.6 kilotorr) to 70 psi (3.6 kilotorr). The nitrosopyrimidine (111) can be reduced for a time sufficient to allow the desired completion of the i II1~SP~C-~T, la~-l~s SWO 94/19351 PCT/US94/01728 -9reaction, such as from 10 minutes to one week or more, preferably from about 5 to 48 hours.

Suitable acylating reagents include anhydrides, organic acid halides, mixed anhydrides, activated acid esters, organic acids or mixtures thereof of the formula R5COL (IV) wherein R 5 is as defined before, and L represents a leaving group, such as an anhydride, a halide or an activated ester. Representative acylating reagents include acetic acid (HOAc), acetyl chloride, acetyl bromide, acetic anhydride, benzoyl chloride, aryl substituted arylacetic acids or derivatives such as para-trifluorophenylacetyl chloride, para-dimethylaminophenylacetyl chloride and paratrifluorophenyl acid. Other acylating agents can include aryl substituted aryl acetic acid chlorides and anhydrides; or can include acetic acid derivatives of formula 2 described in Route 1. The acylating reagents can be employed in amounts effective to acylate the reduced nitrosopyrimidine, and can range from about 1 to about 10 moles of acylating reagent per mole of reduced nitrosopyrimidine, preferably from about 1 to 4 moles of acylating reagent. Acylation can be carried out at temperatures effective to give the amidopyrimidine and can range from about -40°C to about 50°C, more preferably from about 00 to about 400C. The acylation of the reduced nitrosopyrimidine can be carried out at pressures ranging from about ambient to about 400 pounds per square inch (psi) for a time sufficient to allow the desired completion of the reaction, such as from 10 minutes to 48 hours or more.

Suitable acylating catalysts include dimethylaniline, dimethylaminopyridine (DMAP) and imidazole. The acylating catalysts can be employed in amounts effective to catalyze acylation of the reduced nitrosopyrimidine. Such amounts can range from about 1 to about mole percent acylating catalyst, preferably from about 1 to 10 mole percent.

Suitable coupling reagents include carbodiimides such as dicyclohexylcarbodiimide or 1-(3-dimethylaminopropyl)-3ethylcarbodiimide hydrochloride (DEC). The latter coupling reagent may incorporate its own catalyst(s), such as benztriazoles, imidizoles, N-hydroxysuccinimides, and DMAP. The coupling reagent can be employed in an amount effective to aid acylation of the reduced nitrosopyrimidine. Such amounts can range from about 1 to about moles coupling reagent, preferably from about 1 to 4 moles.

I 9 s~ -~sFI-a I II II I I WO 94/19351 PCT/US94/01728 The reduction and acylation steps can be carried out in a solvent compatible with the reactants. Such solvents include ethers, water, bases, acids, dimethylformamide (DMF) or mixtures thereof. Ethers include diethylether (Et 2 tetrahydrofuran (THF), tertiary butyl methyl ether ((CH 3 3 COCH3) and dimethoxyethane. Bases include hydroxides, carbonates or bicarbonates of alkali and alkaline earth metals such as lithium, sodium, potassium, magnesium, calcium or barium. Preferably the base is in the aqueous form. Acids include compounds of the formula

R

5 COL (IV) wherein R 5 is as defined hereinbefore, such as HOAc, propionic acid, butyric acid, or anhydrides of any of the above acids.

Alternatively, the acids can include acidic acid derivatives of formula 2 described in Route 1. Mixtures of any of the above solvents can be employed. The amount of solvent should be sufficient to provide a mixable slurry of the reactants.

Suitable phase transfer catalysts for promoting acylation in the presence of aqueous or organic media of reduced nitrosopyrimidine include tetra-substituted phosphonium salts, quaternary ammonium salts, such as tetrabutylammonium, tetramethylammonium, benzyltributylammonium, with a complementary counterion such as sulfate, hydroxide or chloride. Suitable solvents for use with the phase transfer catalysts include aliphatic hydrocarbons such as C-5 to C-20 alkanes, including heptane, or aromatic hydrocarbons such as toluene, benzene and xylenes; and chlorinated hydrocarbons such as methylene chloride

(CH

2

CI

2 dichlorethane, chlorobenzene; or ethers such as described hereinbefore.

Amidopyrimidine V, wherein Z is =0 and R 4 and R 5 are both methyl is described in A. Branfman et al., Drug Metabolism and Disposition, (1983), Vol. 11, pp. 206-210 and V.I. Khmelevskii et al. J. Gen.

Chem., U.S.S.R. (1958), Vol. 28, pp. 2016-2020.

The halopurine compound of formula VI can be prepared by reacting an amidopyrimidine with an effective amount of a halogenating/cyclizing reagent, optionally in the presence of one or more additional halide sources, and also optionally in the presence of phase transfer catalysts. Suitable halogenating/cyclizing reagents include phosphorous trihalide, a phosphorus pentahalide, organophosphorous halides, a phosphorus oxyhalide, thionyl halide, sulfuryl halide or mixtures thereof. The halogenating/cyclizing reagent can be employed in amounts ranging from about equimolar to excess moles per mole of L I~ b 3 ise I r I r SWO 94/19351 PCT~US9401728 -11amidopyrimidine, preferably from about one mole to about 50 moles of the halogenating/cyclizing reagent. The reaction can be carried out at temperatures ranging from about ambient to the boiling point of the solvent(s) employed, more preferably from about 500C to about 1500C.

Also preferred is that the reactants are contacted at ambient pressures, although pressures greater than ambient can be employed. The reactants can be reacted for a time sufficient to allow the desired completion of the reaction, such as from 10 minutes to about one week or more.

Suitable additional halide sources include ammonium halide such as NH 4 CI, NH 4 Br or NH 4 1, alkali metal halides such as LiCI, LiBr or Lil, halogen gases such as chlorine, bromine or iodine, and hydrogen halides such as HCI, HBr or HI. Such halide sources, together with the halogenating/cyclizing reagent, can facilitate conversion of amidopyrimidine to halopurine The halide sources can be employed in amounts ranging from about 0.1 moles to excess moles per mole of amidopyrimidine preferably from about 0.1 mole to about 5 moles.

Suitable phase transfer catalysts for converting amidopyrimidine to halopurine (VI) can include those described for the above conversion of the reduced nitrosopyrimidine to amidopyrimidine. Such amounts can range from about 0.1 to about moles of phase transfer catalyst per mole of amidopyrimidine, preferably from about 0.1 to about 5 moles.

The process for preparing halopurine VI can be carried out neat or with any solvent compatible with the reactants. Suitable solvents include aliphatic hydrocarbons such as C-5 to C-20 alkanes, preferably heptane, aromatic hydrocarbons such as toluene, benzene or xylenes, chlorinated hydrocarbons such as CH 2 CI2, dichlorethane or chlorobenzenes, or mixtures of any of the above. Where the process is conducted neat, an excess amount of the halogenating/cyclizing reagent can be employed. The amount of solvent should be sufficient to provide a mixable slurry of the reactants.

The substituted aminopurine of formula VIII can be prepared by reacting halopurine VI with a suitable amine, optionally in the presence of an added base, optionally in the presence of phase transfer catalysts such as those described for promoting acylation.

Suitable amines can be those as described in PCT/US91/04154 of the formula: ~I rsas Ipl I_ s I I -I I I I WO 94/19351 PCT/US94/01728 -12-

H

2

N

(CH2)n-OH Ral Rbl R c 1

R

dl

VII

wherein Ral, Rbl, R c l Rdl and n are as defined above. Alternatively, the salts of amines can be employed, such as the HCI salt. The amine can be employed in amounts effective to give compound VIII, and can range from about equimolar to about 20 moles per mole of halopurine VI, preferably from about one to about four moles of amine. Suitable added bases can be either organic, inorganic or mixtures thereof. Organic basee can include nitrogen containing bases such as N-methylpyrrolidinone (NMP), triethylamine (Et 3 diisopropylethylamine (iPr 2 NEt), aniline, pyridine, 1,8-bis(dimethylamino)napthalene, polyvinyl pyridine, DMAP, dimethylaniline, leutidine, sodium tertiary butoxide and the like. It should be noted that the amine reactants described in PCT/US91/04154 can also be employed as a base. Suitable inorganic bases can include hydroxide, carbonates and bicarbonates of alkali and alkaline earth metals of Groups IA and IIA of the periodic table. Such bases can include NaOH, KOH, and sodium and potassium carbonates. The process can be carried out at temperatures effective to give the substituted aminopurine (VII), and can range from about -20°C to 200°C, more preferably from about ambient to about 1500C. Also preferred is that the reactants are contacted at ambient pressures, although pressures greater than ambient can be employed. Optionally, the process can be carried out in a solvent compatible with the reactants. Such solvents can include any of the organic bases described above, acetonitrile (CHsCN), ethers such as THF, (CH 3 3

COCH

3 dimethoxyethane, amides such as DMF and acetamide, aliphatic hydrocarbons such as C-5 to C-20 alkanes and chlorinated hydrocarbons as described above, or mixtures of any of the above solvents. The amount of solvent should be sufficient to provide a mixable slurry of the reactants. Optionally, the process can be carried out with phase-transfer agents, such as those defined hereinbefore.

The reactants can be contacted for a time sufficient to allow the desired completion of the reaction, such as from one to 96 hours or more, preferably from about 24 to about 72 hours.

The desired polycyclic guanine (II) can be prepared by known methods, such as those described in PCT/US91/04154. Generally, a substituted aminopurine Vll can be converted to polycyclic guanine XI L~I _I L~ 'I -h _I L ,WO 94/19351 PCT/US94/01728 -13by ring closure with a suitable dehydrating agent such as thionyl chloride

(SOCI

2 or triphenylphosphine dibromide according to known procedures or procedures analogous to known procedures.

Recovery of compounds II, V, VI and VIII from the reaction mixture can be made using conventional recovery procedures, such as by extraction, crystallization, filtration and/or removal of any solvents present.

In addition to using the novel process described above, the compounds of the present invention can be prepared by several routes as 'escribed hereinafter. Variations of these routes can be employed, as well as other routes known to those skilled in the art, such as those described in W091/19717, incorporated herein by reference.

For compounds of formula Ia wherein R a is hydrogen and Rb and RC, together with the carbon atoms to which they are attached, form a saturated ring of 5 carbons, the following routes 1-3 can be used: Route 1 0 DEC, DMAP, DMF R1 Me, NHCH 2 Ph Me., N~

R

I I

R

ORH 2 HOOCN J R2

H

1 2 R3

H

POl POC3 Me.R

R

2 OH

MR

H

2 N NMP, iPr 2 NEt r> R2 1 S, WO 4/19351 PCT/US94/01728 -14-

SOC

2 o 0

R,

6 Me, N1 NN R3 7 0 H Ri

MI

Pd(OH) 2 H2 or 10% Pd/C NH 4 00CH N.

CH

3 OH, Reflux H Ila Route 1, wherein R 1

R

2 and R 3 are as defined above, involves coupling a uracil derivative of formula 1 with an acetic acid derivative of formula 2 using standard peptide coupling techniques, e.g.

using a coupling agent such as DEC and an activating agent such as DMAP in a suitable inert solvent such as DMF. The substituted uracil derivative of formula 3 is then treated with a halide-forming agent such as POCl3, and the resultant compound of formula 4 is aminated by reacting with trans-2-hydroxycyclopentylamine (formula 5) at elevated temperatures (100-1500C) in the presence of i-Pr 2 NEt in a solvent such as NMP. The resultant compound of formula 6 is then treated with a dehydrating agent such as SOC12 to form the polycyclic ring structure of formula Z. Compounds of formula I can then be obtained by removing the amino-protecting benzyl group, e.g. by hydrogenation with a suitable palladium catalyst and hydrogen or HCO 2

NH

4 Starting uracil derivatives of formula 1 can be prepared by methods known in the art, for example by methods disclosed in W091/19717. The aminoalcohol of formula 5 can be prepared by methods known in the art and by the following procedure:

OH

BnNH2 BnHN A H2, Pd/C Reflux wherein Bn is benzyl. In the reaction scheme, cyclopentene oxide is converted to the corresponding N-benzyl trans-hydroxycycloalkylamine of formula 9 by refluxing with benzyl amine, and the benzyl protecting group slL -I II S WO 94/19351 PCT/US94/01728 is the removed, e.g. by hydrogenation, to obtain the transhydroxycycloalkylamine of formula Route 2 Me 1. LDA/THF MeNN

R

Pd(OH) 2

H

2 1HOC R a i Hr IQ 11 n R3 12 Pd(OH)2, H2 12 -HCI Ia In Route 2, the known starting tetracycle of formula 10 is reacted with a base such as lithiumdiisopropylamide (LDA) in a suitable solvent such as THF, then with a benzaldehyde of formula 11. The resultant compound of formula 12 is then reduced, for example by hydrogenation with palladium catalyst and hydrogen in the presence of HCI to obtain a compound of formula Ia.

Route 3 0 H O H Me, Me, N M N J Br 2 NaOAc, HOAc Br N 50 0 C N N Hi'" IH Hi" l H 3 1d 14

THF

Ar Br Ar ZnBr Zn 16 Pd(PPh 3 4 PPh 3

NMP

14 16 .Ia "Ia 100oo In Route 3, the known starting material of formula J3 is brominated, for example by reaction with a mixture of bromine and sodium acetate in HOAc. The reagent of formula 16 is prepared by treating with zinc a bromomethyl aryl compound of formula 15. (wherein Ar is optionally substituted phenyl or naphthyl). The reagent of formula l.is then reacted with the compound of formula 14 in the presence of a catalyst such as ls Y II

I

WO 94/19351 PCT/US94/01728 -16tetrakis-(triphenylphosphine)palladium (Pd(PPh 3 4 and triphenylphosphine (PPh 3 in the presence of a polar solvent such as NMP under an inert atmosphere.

Other compounds of formula I can be prepared by the following routes 4-6: Route 4 EtOOC.

NH R CH 3 NCO, EtOOC

H

2 NZ =CH= 2 pyridine NH CH MeHNCOHN CH R2 17 R3 Me, 0 SNaO NH N CH2 R 2 1

R

3 Me, 0 NJ R 2 19 POCa T ClN CHc, r y VIa R3 Via I Route 4, wherein R 1

R

2 and R 3 are as defined above, involves reacting a substituted imidazole carboxylate of formula 17 with

CH

3 NCO in the presence of a base such as pyridine. The resultant compound of formula 18 is then heated with an aqueous base such as NaOH to form the bicyclic compound of formula 19, which is then treated with a halide-forming agent such as POCI 3 to obtain a compound of formula Via a compound of formula VI wherein R 4 is methyl, X is chloro and R 5 is optionally substituted benzyl). Compounds of formula VIa are treated as described in the claimed process to obtain compounds of formula I.

dl ~LL I-L~L, ~I~I~LsLY I I I I WO 94/19351 PCT/US94/01728 -17- Route Ra NH 2

OH

Rb RC

VII

o r

H

3 C N base CI N

N

0 H3C%

N

>H

HN N N

R

F O H Rb 2

R

c =2- 21 I The first step in Route 5 involves reacting an amino alcohol of formula VII with a chloropurine of formula 20 under the same conditions as step c of the claimed process to obtain a compound of formula 21. Using procedures described in Routes 1 and 2, compounds of formula I can be dehydrated with SOCI 2 or triphenylphosphine dibromide to form a polycyclic compound, which is then reacted with a base such as LDA in a dry solvent such as THF at low temperature, treated with an electrophile such as an aryl aldehyde of formula 11 and hydrogenated to give compounds of formula I.

Compounds of formula Ib wherein Ra is hydrogen and Rb and Rc complete a tetrahydrofuran ring are prepared by the following procedure: Route 6

H

2 N OH R1) Et 3

N

R

3 2 2 2) CH 3

SO

2

CI,

Et 3

N

R:

N

NH

4

HCO

2 23 Pd Pd/C

R,

0 R2 H3CN

R

N N ,IJ H Ib

O

In the above procedure, a chloropurine of formula 4 is reacted at elevated temperature with an amino alcoho' of formula 22 in the presence of a base such as Et 3 N and in a solvent such as NMP, and the ~c I ~I P"L I~F CI I I -r Iq I i 1 I WO 94/19351 PCT/US94/01728 18resultant intermediate product is then reacted with CH 3

SO

2 CI in the presence of a base such as Et 3 N to obtain an intermediate of formula 23.

The intermediate of formula 23 is then subjected to hydrogenolysis, for example by treatment with NH 4

HCO

2 and a palladium catalyst, to obtain a compound of formula Ib.

Intermediate amino alcohols of formula 22 can be prepared by methods known in the art, for example by the following procedure: /-CPBA CH(CH 3

)NH

2 NH OH m-CPBA A O 1 0 0:, o 24 S NH 4

HCO

2 Pd/C wherein 1,4-dihydrofuran is reacted with m-chloroperoxybenzoic acid (m-CPBA) to obtain the compound of formula 24, which is then reacted with R-(+)-a-methylbenzy!amine to give the substituted amino alcohol of formula 25. The diastereomeric intermediates of formula 25 are separated at this stage by recrystallization from CH 2

CI

2 and hexane. Compound is then hydrogenated by refluxing with NH 4

HCO

2 in CH 3 OH over Pd/C to give the trans-substituted aminoalcohol of formula 22.

Starting materials of formulae 1, 2, 5; 10, 11, 1, 15 and 1 are readily available Jr can be prepared by methods well known in the art.

The following examples are presented to illustrate typical intermediate compounds of the present invention, but the scope of the invention is not to be considered limited to such examples. In the formulae, Me is methyl.

Preparation 1 N-(6-Amino-1,2,3,4-tetrahydro-3-methyl-2,4-dioxo-5-pyrimidinyl) acetamide 0 0

CH

3 N. NO CH3 N NHCOCH 3 O N NH2 O N NH 2 H H Method 1: Hydrogenate overnight, in a parr shaker, a mixture of 100 g of 6-amino-3-methyl-5-nitroso-2,4(1H, 3H)-pyrimidinedione and 20 g of i ~L~S g 111 1 g pp I i la~c~ J$P) II I WO 94/19351 PCTfS94/01728 -19- Fcd/C catalyst in 1.0 L of glacial HOAc. Filter the reaction mixture through g of a diatomaceous earth known as celite®, trademark of the Johns- Manville Products Corporation, Celite Division, Manville, NJ. Wash the celite cake with 125 mL of glacial HOAc and save the cake. Add 45 mL of acetic anhydride along with optionally, a trace of DMAP as a catalyst to the filtrate. Stir the mixture overnight at 400C and then concentrate to dryness under vacuum. Add 125 mL of water to the resultant solid and stir the mixture at 0°C in an ice-water bath for 1 hour. Filter the resultant solid and dry in a draft oven at 400- 500C to obtain 26.3 g of product. Extract the celite cake with eight 250 mL portions of hot HOAc. Concentrate the extracts to obtain a tan solid, slurry in 300 mL water and then treat as described above to give 85.4 g of product. Combining the solids gives 111.7 g (95% crude yield) of the title compound. Recrystallize a sample from hot CH30H/HOAc!C to give the purified title compound as a white solid, melting point greater than 250°C. MS 198 Method 2: Hydrogenate overnight, in a parr shaker, a suspension of 1 g of 6-amino-3-methyl-5-nitroso-2,4(1H, 3H)-pyrimidinedione and 0.1g Pd/C catalyst in 12.2 mL of 1 N NaOH. Filter the catalyst, wash with a small amount of 1N NaOH, cool the filtrate to 0°C, stir and add 2 mL of acetic anhydride. Stir for two hours while allowing the reaction mixture to warm to room temperature, filter, wash with water followed by hexanes and dry overnight in a draft oven at 450C to obtain 0.82g (70% yield of title compound, a light tan solid.

Preparation 2 N-(6-Amino-1,2,3,4-tetrahydro-3-methyl-2,4-dioxo-5-pyrimidinyl) phenylacetamide O 0 O N 1 o CHa 3 .N J NO CH 3 .N NH- O N NH 2 O N NH 2 H H In a parr shaker, hydrogenate overnight a suspension of ig of 6-amino-3methyl-5-nitroso-2,4(1H, 3H)-pyrimidinedione and 0.1g of 5% Pd/C catalyst in 6.2 mL of 1N NaOH. Filter the catalyst, wash with a small amount of 1N NaOH, cool the filtrate to 0°C, stir and add 0.75mL of benzoyl chloride. Stir for 1 hr, filter (save the filtrate), wash the product with water followed by hexanes and dry in a draft oven at 450C to obtain 1.06 g of product as a light yellow solid. The filtrate is stirred at 0°C and 0.375 mL benzoyl chloride is added. From this, 0.31g of product is r I I I ~-4s I I IIRI11 _L _L L SWO 94/19351 PCT/US94/01728 collected as above, to give 1.36g (89% total yield).of title compound, MS

(CI/CH

4 261 m.p. 2500C.

Preparation 3 N-(4-Amino-1,6 dihydro-2-methoxy-1-methyl-6-oxo-5pyrimidinyl)acetamide 0 o

CH

3 N O CH 3

NHCOCH

3

CH

3 aO' I NH CH 3 aO NB NH 2 In a parr shaker, hydrogenate overnight a mixture of 4.5 g of 6-amino-2methoxy-3-methyl-5-nitroso-4(3H)-pyrimidinedione and 0.45 g of 5% Pd/C catalyst in 45 mL of glacial HOAc. Filter the reaction mixture through 1.1 g celite. Wash the celite cake with 1.2 mL glacial HOAc. To the filtrate add 2.8 mL acetic anhydride, and optionally, a trace of DMAP as a catalyst.

Stir the mixture 1hour at room temperature and then filter to collect first crop (0.86 g) of product. Concentrate the mother liquor and filter to collect additional 2.5 g of product. Finally, concentrate the mother liquor to obtain 1.2 g of additional product, to give a total of 4.56 g (90%yield) of the title compound a light tan solid. MS: (FAB) 213 Preparation 3.1 N-(6-Amino-1,2,3,4-tetrahydro-3-methyl-2,4-dioxo-5-pyrimidinyl)-4- (trifluoromethyl)benzeneacetamide R NO R'N-

CF

O N NH2 O N NH 2 H H Hydrogenate, in a Parr shaker, a suspension of 2.3g of 6-amino-3-methyl- 5-nitroso-2,4(1H, 3H)-pyrimidinedione and 0.23g 5% Pd/C in 13 ml 1N NaOH overnight. Filter the catalyst, wash with a small amount of 1N NaOH, cool the filtrate to 0°C, stir and add 4.6g of p-trifluoromethyl phenyl acetyl chloride. Stir for two hours while allowing it to attain room temperature, filter, wash with water followed by hexanes, and dry overnight in a draft oven (450C) to obtain 5.2g of the title compound, a light tan solid. Suspend this solid in CH2CI2, stir for 1h, filter and dry to obtain 4.0g of the title compound as an oft-white solid, suitable for further reactions, m. p. >3000 C. MS 342 Cl (CH4): 343 I I ~I -3 Is Le I I II WO 94/19351 PCT/US94/01728 -21 Preparation 4 2-Chloro-1 ,7-dibydro-1,8-dimethyl-6H-purine-6-one O O H

O

CHN NHCOCH 3 CH3 N N CH 3

NHCOCH

3 N I Js CH3 0 N NH 2 Cl N N H 3 CO N NH 2

H

Method 1: Reflux a suspension of 20 g the product of Preparation 1 in 300 mL of POCIl for 2-4 days, until the reaction is completed as determined by thin-layer chromatography (TLC). Remove POCI 3 under reduced pressure and add a small amount of ice cold water to the resultant dark gummy solid. Stir this mixture vigorously and then bring to a neutral pH at 00-50C by a slow addition of ice cold NH 4 0H. Filter the resultant orange-tan solid, wash with a small amount of ice cold water and dry in a draft oven at 400C to give 16.5 g (crude yield 82%) of the title compound. MS (CI/CH4)199:201 in 3:1 ratio m.p. >2500C.

Method 2: Reflux a mixture of 2 g of the product of Preparation 1 and 0.76g of NH 4 CI in 30mL of POCI 3 gently for 1-2 days, until the reaction is completed as determined by TLC. Remove POCIs under reduced pressure and add a small amount of ice cold water to the resultant gum.

Stir this mixture vigorously and then bring to neutral pH at 0°-50C by a slow addition of ice cold NH 4 0H. Filter the resultant orange-tan solid, wash with a small amount of ice cold water and dry in a draft oven at 400C to give 1.7g (crude yield 83.5%) of the title compound.

Method 3: Reflux a suspension of 0.55 g the product of Preparation 3 in 12 mL of POCI 3 gently for 2-4 days, until the reaction is completed as determined by TLC. Remove POCl 3 and neutralize as described in Method 1. Filter the resultant light brown solid product and dry in a draft oven at 40°C to obtain 0.3 g of product. Extract the mother liquor with ethyl acetate, dry the organic layer over anhydrous Na 2

SO

4 and concentrate under reduced pressure to obtain 0.05 g of additional product.

Finally concentrate the mother liquor to dryness, extract with methanol in ethylacetate (EtOAc), filter and remove the solvents to obtain additional 0.56 g of product, to give a total of 0.41g (80%yield) of the title compound.

~--IYL IIY CIC- 1~qes I~Pa s On 0-p IWO 94/19351 PCT/US94/01728 -22- Preparation 4.1 2-Chloro-1,7-dihydro- 1-methyl-8-[[(4-trifluoromethyl)phenyl]methyl]- 6H-purin-6-one 0 H O 0

H

R 'N NH -CFN3

R

O N NH 2 Cl N N H s .CF 3 Heat a mixture of 5g of N-(6-Amino-1,2,3,4-tetrahydro-3-methyl-2,4-dioxo- 5-pyrimidinyl)-(4'-trifluoromethyl)phenylacetamide and 1.25g NH 4 CI in

POCI

3 at 70° for 24h (until consumption of the starting material as judged by tic). Gently reflux (-1100C) the reaction mixture for 3 days.

Remove POCI 3 under reduced pressure. Add a small amount of ice cold CH2CI2 to the resultant gum. Stir this mixture vigorously, then bring to a pH of 9-10 at 0°-50 C by a slow addition of ice cold NH 4 0H. Remove CH2CI2, add a small amount of water (and NH40H if needed to maintain neutral to alkaline pH), filter the resultant dark brown solid, wash with a small amount of ice cold water and dry in a draft oven at 40 0 C to obtain 5.3 g of the title compound. Slurry this in CH2CI2, filter and dry to obtain 4.6 g (90% mass balance) of the title compound, a light brown solid, suitable for further reactions. MS (CI/CH 4 343:345 in 3:1 ratio; El 342:344 in 3:1 ratio.

Preparation 1,7-Dihydro-2-[(2R-hydroxy-R-cyclopentyl)amino]-I ,8-dimethyl- 6H-purin-6-one 13N NX N CI N N HN NI-N b HOH Reflux a mixture of 12.5 g of the product of Preparation 4, 8 g of 2Rhydroxy-R-cyclopentyl amine, and 31.5 mL of Et 3 N in 83 mL of CH3CN for 2-3 days, until the reactioni is completed as determined by TLC. Remove the volatiles in the reaction mixture under vacuum and then treat with ice cold water. Stir this suspension at 0-5°C and filter. Wash the solid with ice cold water and dry in a draft oven at 400C to give 14.4 g (crude yield 87%) of the title compound, a light brown solid, m.p. 235-2450C (decomposition) MS (CI/CH 4 264 El 263. This compound can be I- 5 L- I PCji~ WO 94/119351 PCT/US94/01728 -23converted to a 2-methyl-polycyclic guanine derivative by employing ring closure procedures described herein or in WO 91/19717.

Preparation 5.1 1,7-Dihydro-2-[(2R-hydroxy-R-cyclopentyl)amino]-1-methyl-8-[[(4- (trifluoromethyl) phenyl]methyl]-6 H-purin-6-one O H CHH CH 3

N

C N NN _N N CF3

HN

CF3 b H-OH Reflux (-1100C) a mixture of 90g of the product of Preparation 4.1, 36.3g of 2R-hydroxy-R-cyclopentyl amine, and 151 ml iPr 2 NEt (Hunig's base) in 151 ml NMP for 24 h (until complete reaction as judged by tic). Allow to attain room temperature, add 1 L ice cold water, stir vigorously for 1h and then pour this in 3.5 L water. Stir for 18h, filter and dry in a draft oven at to obtain 87 g (81% mass balance) of the title compound, a light tan solid, suitable for further reaction. MS (CI/CH 4 408 El 407 m.p. 2850C (decomposition).

The following exemplify preparation of the 2-benzyl-polycylic guanine derivatives.

Example 1 0 H Me,

N

NA

N

H11 I H Step 1: Dissolve 6-amino-3-methyi-5-(phenylmethylamino)pyrimidine- 2,4-dione (12.3 phenylacetic acid 6.80 DEC (9.55 g) and DMAP g) in dry DMF and stir the reaction mixture overnight at room temperature. Pour the reaction mixture onto ice, filter the product and wash with Et20 to obtain: IP I -q 'g ~Y r Ilpl I WO 94/19351 PTU9112 PCT/US94/01728 24 2 MIS, FAB 365

H

Using appropriate starting materiais and essentially the same procedure, the following compounds can also be prepared: 1.1a H NMR (DM30, 300 MHz) 563.00 (3H, s, M 0, NCH 3 3.48 (2H, bs, COCH 2 4.55 (2H, MN F AB, JA,B=14.00 Hz, NCH 2 6.25 (2H, bs,

NH

2 7.0-7.50 (9H, M, C 6

H

5 and C 6

H

4

H

1.1b 1 HNMR (DM30, 200 MHz) 853.00 (3H, s, NOH3), 3.35 (2k, bs, 0001,2), 3.70 (3H, s, M1 e, 000H 3 4.52 (2H, AB, JA,B=1 5.00 Hz, N~ NCH 2 6.18 (2H, bs, NH 2 6.8 and 7.05 NF (4H, 2d, G 6 H4OMe), 7.2-7.40(5H, m, H 0 6

H

5 1.lc 1 H NMVR (DM30, 300 MHz) 6 3.00 (3H, s, M 0. o NCH 3 3.60 (2H, bs, 000H 2 4.55 (2H, M e N F, AB, JAB1.0Hz,

NCH

2 6.30 (2H, bs, 04 NF NH 2 7.10-7.50 (7H, M, C 6

HI

5 and H

C

6

H

4 0F 3 ),7.70 (2H, d, 0 6

H

4 0F 3 1.1d 1 H NMR (DMSO, 200 MP 62.88 (6H, a, GOJ N(rCI l 3 2 3.05 (3H, S, NCirj), 3.35 (2H, Me, 0 bs, COCH 2 4.58 (2H, AD, JAB=14.OOHZ, N N-J m NCH 2 6.18 (2H, bs, NH 2 6.65 and 6.96 0 N 0 (4H, 2d, J=8.65', C 6

H

4 NMe 2 7.25 (5H, m, H 0 6

H

5 PCT[US94/01728 IWO 94/19351 l.1e 1 H NMR (DMSO, 200 MHz) 5 3.15 (3H, s, j~jJ NCH 3 3.95 (2H, bs, COCH 2 3.60 (3H, s, 0 OMe OCH 3 3.80 (6H, s, 2OCH3), 4.60 (2H, AB, Me-.

N~ JA,B=15.00 Hz, NCH 2 6.18 (2H, bs, 0 ~N ome NH 2 6.50 (2H, S C 6

H

2 (OMe)3), 7.2-7.50 H (5H, m, 0 6 1 .lf 1 H NMR (DMSO, 200 MHz) 8 3.10 (3H, s', Me 0 OMe NCH 3 4.20 (2H, bs, COCH 2 3.70 (6H, s, N;I N 200H 3 4.50 (2H, AB, JA,B1 6.00 Hz, N NFP2NCH 2 6.10 (2H, bs, NH 2 6.80 (3H, m H 0 6

H

3 (OMe) 2 7.2-7.50 (5H, M, 0 6

H

5 31gp 2- Dissolve compound 1.1 (5.0g) in 150 mL of POC1 3 and reflux for 8h. Cool the reaction mixture and pour into 300 mL of hexane. Let the reaction mixture stand for 0.5h. Decant the solvent, cool the remaining residue, adjust to pH 8 with 3N NaOH, extract with WOO0 mL of CH 2 CIg, dry and evaporate the solvent. Column chromatograph the residue (2:98_ MeOH :0H 2 C1 2 to obtain the product: 1.2 .00-N. 1H NMR (ODC1 3 200 MHz) 5 3.80 (3H, s, me,

NCH

3 4.30 (2H, s, OH 2 Ph), 5.50 (2H, s, N

NCH

2 Ph), 7.35 (10H, 20 6

H

5 C I 'N

N

Using appropriate starting materials and essentially the samne procedure, folloWing, cmpounds can also be prepared: 1 .2a 1 .2b 1 H NMR (ODC1 3 200 MHz) 5 3.75 (3H, s,

NOH

3 4.10 (2H, s, OH 2 PhF), 5,50 (2H, Si

NCH

2 6.90-7.20 and 7.30 (9H, 2m,

C

6

H

4 F and 0 6

H

5 1 H NMR (0D01 3 200 MHz) 8 3.70 (3H, s,

NCH

3 3.80 (3H, s, 00H 3 4.05 (2H, s,

OH

2 PhOMe), 5.50 (2H, s, NOH 2 Ph), 6.80 (2H, d, J=8.50 HlZ, C 6

H

4 OMe), 7.0-7,10 and 7.30 (7H, 2m, CrH 4 OMe and 0 6

H

5 -OMe I WO 94/19351 WO 9419351PCT[US94101728 -26 1 .2c M.0 1 .2d 1 .2e Me.

CI

N

1 .2f 0

U

Me. NA C I 1 H NMR klC'DCI 3 200 MHz) 5 3.75 (3H, s,

NCH

3 4.18 (2H, s, CH 2 PhCF 3 5.55 (2H,

CF

3 s, NCH 2 Ph), 7.00 (2H, M, C 6

H

4 0F 3 7.20- 1X 7.40 (5H, M, C 6

H

5 7.50 (2H. d, J=8.20 Hz, C 6

H

4 CF3) 1 H NMR (ODC1 3 200 MHz) 8 2.90 (6H,

N(CH

3 2 3.70 (3H, s, NCH 3 4.05 (2H, s, NMe2 CH 2 PhNMe2), 5.45(2H, s, NCH 2 Ph), r (2H, d, O 6

H

4 NMe 2 7.0-7.10 and 7.30 (7H, M, C 6

H

4 NMe 2 and C 6

H

5 1H NMR (ODC1 3 200 MHz) 5 3.35 (3H, s,

NCH

3 3.60 (3H, s, OCH3), 3.65 (6H, s, vle 200H 3 4.15 (2H, s, CH2Ph(OMe)3), 5.70 OMe (2H, s, NCH 2 Ph), 6.45 (2H, s, C 6

H

2 OMe (OMe)3), 7.10 and 7.30 (5H, 2m, 1 H NMR (ODC1 3 200 MHz) 5 3.72 (3H, s,

NOH

3 3.78 and 3.85 (6H, 2s, 20CH3), -Oe 4.08 (2H, s, CH 2 Ph(OMe) 2 5.50 (2H, s,

NCH

2 Ph), 6.70 (3H, M, C6H 3 7.05 and 7.30 (5H, 2m, C 6

H

5 Step 3: Suspend compound 1.2 (2.70 trans-2-hydroxycyclopentylamine (1.40g) and i-Pr 2 NEt (5.20 mL) in 10 mL of NMP and seal the reaction vessel. Keep the mixture at 120-1250C for 6-8h. Cool the reaction mixture, add 20 mL of ice water and filter the precipitate. Wash the precipitate with cold water and,,dry to obtain the product: 1.3 Me, 0 1 HNMR (ODC1 3 200 MHz) 5 1.40-2.20

N

HN N N CH 2 Ph), 4.10 (2H, in), 5.50 (2H, AB, I %H JA,B=14.0 Hz, NCH 2 Ph), 7.20 -7.30 -Ir M, C 6

H

5 Using appropriate starting materials and essentially the same procedure, the following compounds can also be prepared: WO 94/19351 PTU9/12 PCTIUS94/01728 -27 1 .3a Me 0 HN

Y

6,

OH

1 .3b 1 H NMR (00013, 200 MHz) 8 1.40-2.30 (6H, in), 3.40 (3H, s, NCH 3 4.00 (2H, s,

CH

2 PhF), 4.10 (2H, in), 5.40 (2H, AB, JA,B=1 4.0 Hz,. NCH 2 Ph), 7.20 and 7.30 (9H, 2m, 0 6

H

5 and 0 6

H

4

F)

1 H NMR (00013, 200 MHz) 8 1.45-2.30 (6H, in), 3.40 (3H, s, NCH 3 3.75 (3H, s, M1e 00H 3 4.00 (2H, s, CH 2 PhOMe), 4.10 (2H, mn), 4.75 (1 H, d, J=1.80 Hz), 5.45 (2H, AB, JA,B=1 4.0 Hz, NCH 2 Ph), 6.88 and 7.05 (4H, 2d, J=8.0 Hz, C 6

H

4 OMe), 7.05 and 7.30 (5H, 2m, C 6

H

5 1 .3c ~1.3d 1 H NMR (OrC1 3 300 MHz) 8 1.50-2. (6H, in), 3.48 (3H, s, NCH 3 4.15 (2H, s,

CH

2 PhCF 3 4.10 (2H, in), 4.68 (1 H, d, J=1.80 Hz), 5.50 (2H, AB, JA.,9314.0 Hz,

NCH

2 Ph), 7.05 (2H, dd, J=8.0 and 1.80 Hz, 0 6

H

4 0F 3 7.27 (5H, Mn, 0 6 Hs), 7.51 (2H, d, J=8.0 Hz, C 6

H

4 0F 3 1 H N MR (C D C1 3 200 M Hz) 8 1. 40-2.40 (6H, in), 2.90 (6H, s, N(CH 3 2 3.40 (3H, s,

NCH

3 3.95 (2H, s, 0H 2 PhNMe 2 4.05 (2H, in), 4.80 (1 H, d, J=2.0 Hz). 5.45 (2H, AB, JAB=1 5.0 Hz, NCH 2 Ph), 6.65 and 7.05 (4H, 2d, J=8.0 Hz, C 6

H

4 NMe 2 7.10 and 7.30 (5H, 2m, 0 6

H

5 1 H NMR (00013, 200 MHz) 8 1.45-2.20 (6H, in), 3.40 (3H, s, NCH3), 3.75 (6H, s, 200H 3 3.85 (3H, s, 00H3), 4.00 (2H, s, 0H 2 Ph(OMe) 3 4.10 in), 5.50 (2H, AB, JA,1=4.50 Hz, NCH 2 Ph), 6.35(2H, s, 0 6

H

4 (OMe) 3 7.05 and 7.30 (5H, 2m, 0 6 1 .3e 0 OMe HN. OMe e IWO 94/19351 PTU9/12 PCTIUS94/01728 -28 1 .3f 1 H NMR (ODC1 3 200 MHz) 8 1.45-2.30 0 O~e (6H, in), 3.45 (3H, s, NCH 3 3.78 and 3.85 0OMe (6H, 2s, 200H 3 4.05 (2H, s, CH 2 Ph- (OMe)2),4.10 (2H, in), 5.45 (2H, AB3,, HN N NJA,B=15.0 Hz, NCH 2 Ph), 6.75 (3H, 2d, 110H C 6

H

4 (OMe) 2 7.05 and 7.28 (5H, 2m, 6 H 06H5) Step 4: Add 2.30 g Of SOC1 2 (19.35 mmol) to a solution of compound 1.3 (2.75 g, 6.45 mmol) in CH 2

CI

2 and stir the reaction mixture overnight.

Wash the reaction mixture with cold 2N NaOH, dry and evaporate the solvent. Chromatograph the residue on silica, eluting with CH 2

CI

2

/CH

3

OH

(98:2) to give the product: 1.4 1 H NMR (0D01 3 200 MHz) 8 1.45-2.05 Me r (5H, in), 2.20 (1 H, dd), 3.40 (3H, s, NCHs), N 3.98 (2H, s, CH 2 Ph), 4.75 (1 H, t) 4.90 (1 H, NNN 5.50 (2H, a, NCH 2 Ph), 7.10 and 7.30 Hi11 2 &(iiH (10H, 2mn, C 6

H

5 Using appropriate starting materials and essentially the same procedure, the following compounds can also be prepared: 1.4a

NN

1 .4b 1H NMR (ODC1 3 200 MHz) 8 1.50-2.10 in), 2.30 (1 H, dd), 3.40 (3H, s,

NCH

3 4.00 (2H, s, CH 2 PhF), 4.80 (1 H, t) 4.92 (1 H, 5.42 (2H, s, NCH 2 Ph), 7.10 and 7.32 19H, 2m, C 6

H

5 and 0 6

H

4

F)

1 H NMR (ODC1 3 200 MHz) 8 1.50-2.00 H, in), 2.28 (1 H, dd), 3.35 (3H, s,

NCH

3 3.80 (3H, s, OCH 3 3.01 (2H, s,

CH

2 PhOMe), 4.75 (1H, t) 4.85 t), 5.35 (2H, s, NCH 2 Ph), 6.80 and 7.05 (4H, 2d, J=8.0 Hz, C 6 H4OMe), 7.10 and 7.30 (5H, 2m, GsH 5 Me, OMe

N/

Hils d4N I.WO 94/19351 PTU9/12 PCTIUS94/01728 29 1 .4c 1 H NMR (ODC1 3 200 MHz) 8 1.50-2.10 in), 2.28 (1 H, dd), 3.38 (3H, s, Me. CF3 NCH 3 4.08 (2H, s, CH 2 PhCF 3 4.78 MeN k N .C (1 H, 4.90 (1 H, 5.40 (2H, s, b N NNCH 2 Ph), 7.05 (2H, dd, J=8.0 and 1.80 H" IH Hz, C 6

H

5 7.30 (3H, M, C 6 H5), 7.20 and 7.50 (4H, 2d, J=8.0 Hz, 0 6

H

4 0F 3 1 .4d 1 H NMR (ODC1 3 200 MHz) 5 1.50-2.00 in), 2.30 (1 H, dd), 2.90 (6H, s, Me, 0 NMe 2 N(0H 3 21 (3H, s, NCH 3 3.92 (2H, N N s, CH 2 PhNMe 2 4.72 (1 H, 4.88 (1 H, t), N N N5.35 (2H, s, NCH 2 Ph), 6.65 and 7.00 Hilt I-IH (4H, 2d, J=8.0 Hz, C 6

H

4 NMe 2 7.12 and 7.30 (5H, 2m, C 6 1 .4e 1 H NMR (ODC1 3 200 MHz) 8 1.50-2.00 o Ph OMe (5H, in), 2.25 (I H, dd), 3.35 (3H, s, Me,. N r..,OMe NCH 3 3.70 (6H, s, 200H 3 3.80 (3H, S, I OMe OCH 3 3.95 (2H, s, CH 2 Ph(OMe) 3 4.75' N~ r~ N (1 H, t) 4.90 (1 H, 5.40 (2H, s, NOH 2 Ph), Hit- i H6.30 (2H, S, C 6

H

4 (OMe) 3 7.10 and 7.30 2m, 0 6 1 .4f 0 rPh m Me,. N r OOMe H~r 1 H N MR (ODC1 3 200 MHz) 1.50-2.00 (5H, in), 2.40 (1 H, dd), 3.40 (3H, s, NCI~a), 3.78 (6H.s. 200H 3 3.85 (3H, s,

OCH

3 4.10 (2H, s, CH 2 Ph(OMe) 2 4.75 (1 H, t) 4.85 (1 H, 5.50 (2H, s, NCH 2 Ph), 6.80 and 6.62 (3H, 2m, C6H 4 (OMe) 2 7.10 and 7.30 (5H, 2m, 0 6

H

5 compound 1.4 (4.00 g) and 10 PdIC (4.00 g) in 100 mL of CH 3 OH, add NH 4

HCO

2 and reflux for 6h. Cool the reaction mixture, filter and evaporate the solvent. Add 20 mL of water to the resultant residue, adjust to pH to 8, extract with CH 2

CI

2 dry and evaporate the solvent to produce bIe title compound 1 H NMR (CDC1 3 +drop of

CD

3 OD, 200 MHz) B 1.50 (1 H, in), 1.65 -2.00(5H, in), 2.28 (1 H, J=7.50 12.50 Hz), 3.28 (3H, s, NCH3), 4.18 (2H, s, CH 2 Ph), 4.75(1 H, t, J=7.00 Hz), 4.90 (1 H, t, J=7.00 Hz), 7.30 (5H, Mn, C 6

H

5

W

I WO 94119351 WO 9419351PCT/1JS94101728 Using the same procedure, hydrogenate compounds 1 .4a to 1 .4c of Step 4 to obtain the following compounds: 1A 1 HNMR (CDCI3+drop of CD3QD, 200 0 H MHz) 8 1.50-2.00 (5H, in), 2.25 (1 H, dd, F J=13.00 and 5.00 Hz), 3.38 (3H, s, NOHa), N ~4.15 (2H, S, CH 2

C

6

H

4 4.72 (1 H, t, H J=7.00 Hz), 4.85 (1 H, t, J=7.00 HZ), 6.95 and 7.30 (4H, 2d, J= 10 Hz, C6H 4

F)

1lB 1 H NMR (CDCI3+drop of CD 3 OD, 300 M e HO~ MHz) 8 1.50-2.00 (5H, in), 2.28 (1 H, dd, N N J=1 3.50 and 5.20 Hz), 3.38 (3H, s, NOHa), N N 'N 3.50 (3H, s, OCH 3 4.10 (2H, s, HI1 I iH CH 2

C

6

H

4 OMe), 4.72 (1H, t, J=7.00 Hz), 4.88 (1 H, t, J=7.00 Hz), 6.85 and 7.25 (4H, 2d,,1=10.0 Hz, C6H 4 OMe) 1 H NMR (CDCI 3 +drop of CD 3 OD, 300 0 CF MHz) 8 1.50-2.00 (5H, in), 2.25 (1 H, dd, N N J=1 3.50 and 5.20 Hz), 3.35 (3H, s, NCH3), NO N[N 4.22 (2H, s, CH 2 C6H 4

CF

3 4.72 (1 H, t, H1 I H J=7.00 Hz), 4.88 (1 H, t, J=7.00 Hz), 7.48 and 7.58 2d, J=8.0 Hz, C 6

H

4

CF

3 Example 2 0 H M e I NMe 2 N

N

H" e 'd IH Dissolve compound 1.4d (2.0g) in 100 mL of absolute EtOH, add 5 mL of EtOH saturated with HOI gas and 20% Pd(OH) 2 Hydrogenate the reaction mixture at 60 psi for 24 h. Basify the reaction with NH 4 OH, filter and evaporate the solvent. Redissolve the residue in

CH

2 01 2 wash with water, -ct.rv and evaporate the solvent. Column chromatograph the residue (95:5 0H 2 C1 2 :MeOH) to obtain the title compound: 1 HNMR (CDCI 3 +drop of CD 3 OD, 300 MHz) 8 1.50-2.00 m 2.25 (1 H, dd, J=1 3.50 and 5.20 Hz), 2.95 (6H, s, N(CH 3 2 3.35 (3H, s, NCH 3 4.05 (2H, s, CH 2

C

6

H

4 NMe 2 4.72 (1IH, t, J=7.00 Hz), 4.88 (1 H, t, J=7.00 Hz), 6.70 and 7.15 (4H, 2d, J=8.0 Hz, C 6

H

4 NMe 2 WO 94/19351 PCTIUS94/01728 Using the same procedure, treat the compounds 1.4e and -1.4f to obtain compounds 2A and 2B3, respectively: 2A 1 H NMR (CDCI3+drop of 0D 3 01D, 300 0 H Me MHz) 8 1.50-2.00 (5H, m 2.25 (1 H, ddl, Me, N J=13.50 and 5.20 Hz), 3.40 (3H, s, NCH 3 b OMe 3.85 (9H, s, 300H 3 4.10 (2H, s, H" l'HCH 2

C

6 H4OMe), 4.72 (1 H, t, J=7.00 Hz), 4.88 (1 H, t, J=7.00 Hz), 6.50 (2H, s, C6H2(Ole)3) 28 1 H NMR (CDCI3+drop of CD 3 O1D, 200 0 H Me MHz) 8 1.50-2.00 (5H, in), 2.25 (1 H, dd, Me, N )k-f N J=13.00 and 5.20 Hz), 3.35 (3H, s, NCH 3 N'INN 3.85 and 3.88 (6H, 2s, 200H3), 4.10 (2H, H s, CH 2 C6H4(OMe) 2 4.75 (1 H, t, J=7.00 Hz), 4.85 (1 H, t, J=7.00 Hz), 6.85 (3H,m 3 (OMe) 2 Example 3 0 H Me-, IOH NN H" sd IH Generate LIDA (1.2 mmol) in THF (15 mL) and cool to -780C. Add a THF solution of 7,8,9,9a-hexahydro-5-m ethyl-3- (phe nylm ethyl)cyclopent[4,5]iinidazo[,1,-bjpurin-4-one (1.0 mmol) and stir the reaction for 0.5h at -780C. Add a solution of 4-benzyloxybenzaldehyde (1.0 inmol) in THF, slowly warm the reaction mixture to -400C and stir for 0.5h at -400C. Quench the reaction with a saturated NH 4 CI solution and evaporate the solvent. Proceed as described in Example 2 to obtain the title compound: 1 H NMR (CDCI 3 +drop of CD 3 OD, 300 MHz) 8 1.60 (1 H, in), 1.70-2.00 (4H, in 2.20 (1 H, dd, J=1 0.00 and 4.00 Hz), 3.30 (3H, s,

NCH

3 4.20 (2H, s, CH 2

C

6

H

4 OH), 4.72 (1 H, t, J=7.00 Hz), 4.88 (1 H, t, J=7.00 Hz), 6.75 and 7.12 (4H, 2d, J=8.0 Hz, C 6

H

4

OH).

Using appropriate starting materials and essentially the same procedure, the following compounds can also be prepared: PCTIUS94/01728 IWO 94/19351 32 3A Me, CH 3 N

N

Hio o ml 1 H NMR (CDCI 3 +drop of CD 3 OD, 200 MHz) 8 1.60-2.00 (5H, m 2.28 (1 H, dd J=1 0.00 cand 5.00 Hz), 2.32 (3H, s, 0 6

H

4

CH

3 3.35 (VH, s, NCH3), 4.12 (2H, s, CH 2

C

6

H

4 Me), 4.72 (1 H, t, J=7.00 Hz), 4.88 (1 H, t, J=7.00 Hz), 7.10 and 7.20 (4H, 2d, J=8.0 Hz, C 6

H

4 0H 3 1 H NMR (CDCI3+drop of CD 3 OD, 200 MHz) 81.50-2.00 (5H, 2.28 (1H, dd, J=1 0.00 and 5.00 Hz), 3.35 (3H, s, NCH 3 4.18 (2H, s, CH 2

C

6

H

4 4.72 (1H, t, J=7.00 Hz), 4.88 (1 H, t, J=7.00 Hz), 7.15 and 7.40 (4H, 2d, J=8.0 Hz, 0 6

H

4 0CF: 3 1 H.NMR (300 MHz, ODC1 3 8 1.47-2.31 (in, 6H), 3.05-3.15 (in, 4H), 3.38 3H), 3.78-3.86 (in, 4H), 4.08 2H), 4.73 1H., J=7 Hz), 4.88 1 H, J=7 Hz), 6.84 2H, J=6.7 Hz), 7.24 2H, Hz); 2 +790 (c 0.7, EtOH);

D

Me, N N NN Hil-d 11H 0 3D 1 H.NMR (200 MHz, ODC1 3 8 1.57-2.35 I (in, 1 OH), 3.21-3.29 (in, 4H), 3.40 3H), Me, N N4.05 2H), 4.75 1 H, J=6.9 Hz), 4.91 (t, ~N<UNQ 1H, J=7.0 Hz), 6.50 2H, J=8.4 Hz), 7.14 N" 'I H 2H, J=8.4 Hz); Cl MS 391 1,00%) 3E o H 22.5 (t 1 [a D= 134.30 EtH Me,.N OMe D HN ~I Hy WO 94/19351 PTU9112 Fr-TIUS94/01728 33 o H N N mp 259-2600C; FAB MS 390 100%) H""s- Me, N N .0 H" 'N 0 D(I2. 108.40 (EtOH) 3H 0 H FAB MS 398 100%) Me% 'N

N

Example 4 $.tep React (+)-cis-5,6a,7,8,9,9a-hexahydro-5-methyl-3-(phenylmethyl)cyclopent[4,5jimidazo[2,1-b]purin-4-one with LIDA, then with 4-(1H-pyrrol- 1-yl)benzaldehyde in a manner similar to that described in Example 3.

5I2 Add sodium pellets (24 mg, 1.0 mmol) to a solution of the product of Step 1 (54 mg, 0. 11 mmol) in anhydrous THE (15 mL) and liquid ammonia (35 ml-) at -78 00. Stir the mixture for 5 min at -78 00, then add solid NH 4 CI to quench the reacton. Allow the liquid ammonia to evaporate at room temperature. Triturate the residue with CH 2

CI

2

/CH

3 -:OH filter, dry over MgSO) 4 and concentrate. Chromatograph the residue, eluting with CHC1 3 /MeOH (95:5) to obtain 9.8 mg (23 of the title compound. Cl MS 387 100%); 1 H-NMR (300 MHz, ODC1 3 d 1.50-2.67 (in, 6 H), WO 94/19351 PCT/US94/01728 34 3.36 3 4.15 2 4.70 1 H, J 6 Hz), 4.87 1 H, J 8 Hz), 6.28-6.30 2 7.00-7.02 2 7.23-7.38 4 H) Example 0 H Me..

N

N N iH I OMe Step. Heat a mixture of the compound of Example 1.4b (4.40 g) and pyridine hydrochloride (5.8 g) to 1700C for 4.5h. Cool to room temperature, add an ice cold solution of saturated NaHCO 3 extract with

CH

2

CI

2 wash the organic layers with brine, dry over MgSO 4 filter and concentrate to yield a solid, FAB MS 428 Step 2: Suspend a portion of the product of Step 1 (0.62 g) in DMF mL), cool to 0°C, add N-phenyl trifluoromethane sulfonimde (0.57 g) and

K

2 CO3 (0.39 Allow the reaction to warm to room temperature overnight, then pour the reaction mixture into ice water (0.7 extract with EtOAc, wash the organic layer with brine, dry over MgSO 4 filter and concentrate to dryness.

Ste: Dissolve a portion of the residue of Step 2 (0.52 g) in dioxane (13 mL), add 4-methoxyphenyltrimethyltin (0.5 g) (prepared from reaction of 4bromoanisole with t-butyl lithium and trimethyltin chloride), LiCI (0.125 g), Pd(PPh 3 4 (0.04 and a catalytic amount of BHT. Heat the reaction mixture to reflux for 15h under a nitrogen atmosphere, cool to room temperature, basify with 10% NH 4 0H, and extract with EtOAc. Wash the organic layer with brine, dry over MgSC4, filter and concentrate to dryness. Chromatograph the residue on silica gel using CH 2 C1 2 /CHsOH (90:10) to give a light yellow solid, El MS (70ev) 517 Step 4: Dissolve the solid from Step 3 (0.36 g) in CH 3 OH (50 mL), add

NH

4

HCO

2 (0.66 g) and 10% Pd/C (0.3g Heat this mixture to reflux for 43h, then add more NH 4

HCO

2 (0.66 g) and continue heating for 8 h, cool to room temperature, filter through celite, concentrate to dryness and partition the residue between 10 NaHCO 3 and CH2Cl 2 Dry the organic layers over MgSO 4 concentrate to dryness and chromatograph the residue on silica gel using CH 2

CI

2 /EtOH/NH 4 0H (90:10:1) to give the title [a2 113.90 (MeOH); compound as a colorless solid: D EV MS (70ev) 427 398 (100%).

I II ,WO 94/19351 PCT/US94/01728 Example 6 H H H3C .N Nr'N Ste. 1: To a solution of cis-5,6a,7,8,9,9a-hexahydro-5-methylcyclopenta- [4,5]-imidazo[2,1-b]purin-4(3H)-one (3.38 g, 14.6 mmol) and NaOAc (1.44 g, 17.5 mmol) in HOAc (70 mL) at room temperature and under N 2 add bromine (0.91 mL, 17.5 mmol). Stir at 500C for 16 h, filter the solids, wash with CHCI 3 and air dry to obtain the 2-bromo derivative as a white solid (3.75 g, 1 H-NMR (400 MHz, DMSO-d 6 8 1.68-2.24 6H), 3.32 (s, 3H), 4.81 1H, J=7 Hz), 5.19 1H, J=7 Hz), 10.18 (brs, 1H); CI MS 232 19.8 310 312 [a]D 100.60 (c 0.63, CHO 3

H).

aiSq: To a solution of zinc dust (1.70 g, 26 mmol) in dry THF at room temperature, under N 2 add 1.2-dibromoethane (0.086 mL, 1.0 mmol) and stir at 650C for 1 min. At 0oC, add, dropwise, a solution of 2bromomethylnaphthalene (4.75 g, 21.5 mmol) in dry THF (11 mL) and stir at 000 for 1 h, then at room temperature for 1 h. Heat a portion of the supernatant (3.3 mL, approx. 5.4 mmol) with the product of Step 1 (0.166 g, 0.535 mmol), Pd(PPh 3 4 (0.062 g, 0.0544 mmol) and PPh 3 (0.028 g, 0.11 mmol) in NMP (2 mL) at 1000C under argon for 20 h. Evaporate the solvent, dissolve the resultant residue in CHC1 3

-CH

3 OH wash with sat'd NaHCO 3 dry over MgSO 4 and concentrate. Separate by flash chromatography, eluting with CHC1 3

-CH

3 OH (97-3) to obtain the title compound (0.079 g, 1 H-NMR (400 MHz, CDC3) 8 1.47-2.28 (m, 6H), 3.30 3H), 4.27-4.35 (AB q, 2H), 4.68 1 H, J=7 Hz), 4.81 1H, J=7 Hz), 7.40-7.80 7H); Cl MS 372 100%); S21.7 [OcD 78.50 (c 0.41, CHC1 3 Example 7 -I ~k 'g CI 'I s WO 94/19351 PCT/US94/01728 -36- In a manner similar to that of Example 6, using appropriate starting materials, prepare the title compound: 1 H-NMR (400 MHz, CDC13) 8 1.49-2.28 6H), 3.36 3H), 4.13 2H), 4.73 1H, J=7 Hz), 4.87 1H, J=7 Hz), 7.26-7.27 4H); CI MS 356 358 [oc 2 101.00 (c 0.41, CHCI 3 Example 8 Me, 0

N

N

N N -CH 2 Ph Step 1: Reflux a solution of ethyl 2-benzyl-4-amino-5-imidazole carboxylate (0.89 g, 3.63 mmol) and CH 3 NCO (8 mL) in pyridine (15 mL) for 2h. Cool the reaction mixture, evaporate the solvent, extract the residue with EtOAc, wash with water, dry and evaporate. Column chromatograph the residue, eluting with 2% MeOH in CH 2 Cl 2 TLC Rf=0.3 MeOH in CH 2

CI

2 MS 303 1 H NMR (CDCIs, 200 MHz) 8 1.32 (3H, t, CH 3

CH

2 2.90 (3H, d, NHCH 3 4.05 (2H, s, CH 2 Ph), 4.30 (2H, q,

OCH

2 Me), 7.35 (5H, m, C 6 .tep 2: Reflux the solution of the product of Step 1 (0.85 g) in 10% (by weight) NaOH (15 mL) for 0.5h. Cool the reaction mixture, adjust the reaction mixture to pH6 with HOAc and filter. Dry the solid at high vacuum overnight. 1 H NM (DMSO, 200 MHz) 53.15 (3H, s, NCH 3 3.95 (2H, s,

CH

2 Ph), 7.28 (5H, m, C 6

H

5 Step 3: Reflux a solution of the product of Step 2 (0.84 g) in POCI 3 mL) for 24h. Cool the reaction mixture, pour into hexane (100 mL) and let stand at room temperature for 0.5h. Decant the solvent, treat the resulting gum with ice cold water, neutralize it with 2N NaOH, extract with CH 2

CI

2 2x100 mL), dry and evaporate the solvent. Column chromatograph the residue, eluting with 5% MeOH in CH 2

CI

2 TLC Rf=0.45 MeOH in

CH

2

CI

2 1 H NMR (CDCI3, 200 MHz) 8 3.80 (3H, s, NCH 3 4.30 (2H, s,

CH

2 Ph), 7.35 (5H, m, C 6 Hs).

Ste~ 4: Suspend the product of Step 3 (0.2 g, 0.73 mmol), 1-amino-1cyclopentane methanol (0.17 g, 1.45 mmol) and (i-Pr 2 NEt) (0.25 mL, 1.45 mmol) in NMP (2 mL) and keep the rec''ion mixture at 1300C for 6h. Pour the reaction mixture on ice/water to precipitate the product. Filter the product and dry it under high vacuum. TLC Rf=0.28 CH 3 0H in II I s~ ~I I~ WO 94/19351 PCTIUS94/01728 37

CH

2

CI

2 MVS 354 Fab; I HNMR (CD013 drop of 00300, 200 MHz) 8 1.60-2.00 (8H, in), 3.45 (311, s, NCH 3 3.80 s, CH 2 OH), 4.26 (2H, s, CH 2 Ph), 7.32 (5H, m, C 6

H

5 Bjpt Treat a suspension of the product of Step 4 (0.08 g, 0.22 mmol) in 0H 2 C1 2 (10 mL) with S012 (0.05 at room temperature overnight.

Dilute the reaction mixture with CH 2

CI

2 wash with cold 1N NaOH, dry and evaporate. Column chromatograph the crude title compound, eluting with MeOH in 0H1 2

CI

2 to obtain the title compound. MS 336 1 H NMR (CDCI 3 drop of CD 3 0D, 200 MHz) 8 1.60-2.00 (811, mn), 3.56 s,

NCH-

3 4.18 bs, CO 2 4.22 s, CH 2 Ph), 7.30 M, C 6

H

5 Example 9 Me~, 0 N Me-j N Me .Ste~p 1: Treat a solution of the product of Step 3 of Ex. 8 (0.16 g, 0.58 minol) and 2-am ino-2-m ethyl propanol 0.13 g, 1.45 minol) according to procedure described in Step 4 of Ex. 8: MS 328 FAB; 1 H NMR (ODC1 3 drop of OD 3 OD, 200 MHz) 8 1.45 (6H, S, (0H 3 2 3.45 (3H, s,

NCH-

3 3.75 s, CH 2 OH), 4.20 (2H, s, CH 2 Ph), 7.30 (5H, M, C 6

H

5 StI22 Treat the product of Step 1 according to the procedure of Step in Ex. 8 to produce the crude title compound. Column chromatograph the crude product, eluting with 7% CH 3 0H in 0H 2 012 to obtain the title compound. TLC Rf=0.60 CH 3 OH-- in CH 2

CI

2 MS 310 1

H

NMR (00013 drop of CD 3 OD, 200 MHz) 8 1.45 (6H, s, (0H 3 2 3.40 s, NOH 3 3.90 (2H, s, CO 2 4.16 s, OH 2 Ph), 7.35 in, 0 6

H

5 Examplr jO Me, 0

N

INH

N -CH 2 Ph

N

Me- Me Stg1 Treat the product of Ex. 8, Step 3 (0.20 g, 0.73 minol) and amnino-3-m ethyl- 1-butan ol (0.15 g, 1.45 iniol) using the pr'ocedure of Ex.

WO 94/19351 PTU9/12 PCTIUS94101728 38 8, Step 4: MS 341; TLC Rf=0.40 (10% MeOH in CH 2

CI

2 1 H NMR (ODCd 3 drop of CD 3 OD, 200 MHz) 8 0.96 and 0.98 (6H, 2d, (CH 3 2

CH),

2.00 (1IH, dt, CH(Me) 2 3.45 (3H, s, NCH 3 3.65 and 3.75 (2H, 2dd, J=l15.00 and 6.5 Hz, CH 2 OH), 4.12 (2H, S, CH 2 Ph), 7.28 (5H, M, C6H 5 atp2 Treat the product of Step 1 (0.11 g, 0.32 mmol) with SOC! 2 (0.14 g, 0.08 mL) as described in Step 5 of Ex. B. Column chromatograph the crude product, eluting with 7% CH 3

OH-CH

2

CI

2 to obtain the title compound. TLC Rf=0.45 (10% CH 3 0H in CH 2

CI

2 MS 324 1 H NMR (ODC1 3 drop of CD 3 OD, 200 MHz) 8 1.00 and 1.05 (6H, 2d,

(CH

3 2 CH), 2.10 (1 H, in), 3.62 (3H, s, NCH 3 4.25 (2H, s, CH 2 Ph), 4.15 (1 H, in), 4.40 (2H, mn), 7.28 (5H, in, C 6

H

5 Example 11 N ul N

~H

Step 1:

H

3 C, N

H

Heat a mixture of 2-chloro-l-methyl-7-(phenylmethy)purin-6-one (1 1am ino-7-hydroxybicyclo[3.3.0]octane hydrochloride (0.65 (i-Pr 2 NEt) (1.25 mL) and NMP (3 inL) in a sealed tube at 1400C for 60h. Coo! the reaction to room temperature and pour into water. Collect the solid by filtration, dissolve it in CH 2

CI

2 and chromatograph on silica gel, eluting with CH 2 01 2

/CH

3 OH (95/5) to give a solid. FAB MS 380 (in+H,100%).

Dissolve a portion of this solid (0.82 g) in CH 2

CI

2 (5 mL), and add SOC1 2 inL. Stir the reaction mixture for 2 h, partition between NaHCO 3 and 0H 2 Cl 2 Wash the organic layer with brine, dry over MgSO4, filter and concentrate to dryness to obtain a solid. El MS (70ev) 361 100%).

WO Q4/19351 PCT/US94/01728 -39- H3C AN O H N J N (jAH Add the product of Step 1 (0.36 g) to a solution of LDA (1.2 mmol) in THF (2 mL) at -780C. Stir the reaction mixture for 0.5h, then add a solution of piperonal (0.18 g) in THF and continue stirring at -780C for 1 h. Quench the reaction with HOAc (0.1 mL) and allow the reaction mixture to warm to room temperature, then partition between EtOAc and NaHCO 3 Wash the organic layer with brine, dry over MgSO4, filter and concentrate to dryness to give a solid. El MS (70ev) 511(M+,100%).

Step 3: Place a mixture of the product of Step 2 (0.37 20% palladium hydroxide on charcoal (0.15 EtOH (75 mL) and conc. HCI (0.6 mL) under a H2 atmosphere (60 psi) for 60,h. Remove the catalyst by filtration through celite, concentrate the filtrate to dryness and partition the residue between NaHCO3 and CH2CI2. Wash the organic L:yer with brine, dry over MgSO4, filter and concentrate to dryness. Chromatograph this residue on silica gel, eluting with CH2C0 2

/CH

3 OH (95/5) to obtain the title compound as a colorless solid, mp 239-240 oC, El HRMS calo for

C

22

H

23

N

5 0 3 405.1801, found 405.1806.

Example 12 0 CF 3 H3CN HIl I H 0 Step 1: A: Add m-CPBA (9.5 g, 0.55 mol) to a solution of 1,4-dihydrofuran (3.8 mL., 0.050 mol) in CH 2 CI2 (100 mL) at room temperature and stir for 16 h.

Filter the solids, wash the filtrate with NaHCO 3 dry over MgSO 4 and concentrate to give a clear oil (2.2 g, H-NMR (300 MHz, CDC 3 8 3.65 2 H, J=10.5 Hz), 3.79 2H), 4.02 J=10.6 Hz).

L~ I ~CI I 119 -1 II ~I 1~ WO 94/19351 PCT/US94/01728 B: Heat R-(+)-a-methylbenzylamine (14 mL, 0.011 mol) with the product of Step A (10 g) and water (2 mL) at 100 oC for 24 h. Cool the reaction mixture and recrystallize the solid twice from CH 2

CI

2 -hexanes (1.2 g).

HPLC showed >91% purity. 1 H-NMR (300 MHz, CDCIs) 8 1.37 3H, J=6.7 Hz), 1.90 (br s, 2H), 3.03-3.07 1H), 3.36 (dd, 1H, J=9.3, 4.1 Hz), 3.66 (dd, 1H, J=9.8, 2.4 Hz), 3.86-3.92 2H), 3.98 (dd, 1H, J=9.8, 4.7 Hz), 4.22-4.25 1H), 7.26-7.37 Q: Heat the product of Step B (750 mg) with NH 4

HCO

2 (82u mg) in refluxing CH 3 OH (30 mL) over Pd/C (700 mg) for 1 h. Cool, filter, and concentrate the mixture (300 mg, [o] 2 5 D -110 (c 0.2, MeOH); 1 H-NMR (300 MHz, CDC13) 8 2.40 (br s, 3H), 3.74-3.96 3H), 4.26-4.34 2H), 4.44-4.48 1H).

Step 2: A: Stir 6-amino-3-methyl-5-(phenylmethyleneamino)pyrimidine-2,4-dione (2.46 4-trifluoromethylphenylacetic acid 5.1 g) and DEC (4.8 g) in dry DMF at room temperature for 4 h. Pour the reaction mixture over ice, filter the solids and wash with Et 2 0 (3.80 g, 1H NMR (DMSO-d 6 300 MHz) 8 3.00 3H), 3.58 2H), 4.51 1H, J=14 Hz), 4.59 1H, J=14 Hz), 6.27 (br s, 2H), 7.22-7.32 5H), 7.38 2H, J=8 Hz), 4.59 2H, J=8 Hz), 10.45 (br s, 1H).

B: Reflux the product of Step 2A (1.8 g) in POCI3 (30 mL) for 16 h. Cool the reaction mixture and dilute with hexanes (600 mL). After 4 h, decant the liquid layer from the oily residue, dissolve the residue in CH 2 Cl 2

CH

3 OH wash with NaHCO 3 dry (MgSO 4 and concentrate.

Purify the residue by flash chromatography, eluting with CHCI 3

-CH

3

OH

(97-3) (700 mg, 1 H NMR (CDC13, 300 MHz) 5 3.72 3H), 4.15 (s, 2H), 5.54 2H), 6.99 2H, J=7.9 Hz), 7.19-7.34 5H), 7.49 2H, J=7.9 Hz).

Step 3: Heat the product of Step 2B (0.68 g, 1.57 mmol), the product of Step 1C (0.28 and Et 3 N (0.9 mL) in 20 mL of NMP at 120 oC for 16 h.

Concentrate the mixture to remove NMP. Dissolve the residue in CH 2

CI

2 wash with NaHCO 3 dry over MgSO 4 and concentrate. Purify the residue by flash chromatography, eluting with CHCI 3

-CH

3 OH (95-5) (320 mg, To a solution of the resultant compound (320 mg) and Et 3

N

(0.6 mL) in CH 2

CI

2 (25 mL), add, dropwise, CH 3

SO

2 CI (0.12 mL) and stir for 16 h. Wash the solution with NaHCO 3 dry (MgSO 4 and concentrate. Purify the resultant residue by flash chromatography, eluting with CHCI 3

-CH

3 OH (97-3) (250 mg, 1 H NMR (CDCIs, 300 MHz) d L f aa e~l I ,I S, WO 94119351 PCT/US94/01728 -41- 3.50 3H), 3.72-3.78 2H), 4.10 2H), 4.29 1H, J=9.2 Hz), 4.47 1H, J=11 Hz), 4.96-5.00 1H), 5.12-5.17 1H), 5.40 1H, J=16 Hz), 5.48 1H, J=16 Hz), 7.02-7.05 2H), 7.15 2H, J=8.3 Hz), 7.26- 7.30 3H), 7.51 2H, J=8.4 Hz).

Step 4: Reflux the product of Step 3 (190 mg), NH 4

HCO

2 (200 mg), and Pd/C (150 mg) in CH30H (25 mL) for 1.5 h. Filter the mixture and wash the solids with CH 2

CI

2 -CH30H Combine the filtrate and the washings, wash with NaHCOs dry (MgS0 4 and concentrate.

Purify the resultant residue by flash chromatography, eluting with CHCI 3

CH

3 OH (95-5) to obtain the title compound (100 mg) as a white solid.

[a]25D=+1250 (c 0.4, EtOH); 1 H NMR (CDCI 3 300 MHz) 8 3.48 3H), 3.69-3.76 2H), 4.19 1H, J=10.5 Hz), 4.24 2H), 4.42 1H, J=1 1.3 Hz), 4.92-4.96 1H), 5.10-5.14 1H), 46 2H, J=8.0 Hz), 7.58 2H, J=8.0 Hz).

Example 13 O H CH3. N N CH3, N HN N N N 'N

)CH

2 0H H -H To a suspension of 1.5 g of the product of Preparation 5 in 45 ml CH 2

CI

2 at room temperature under N 2 add 1.7 ml of SOCI 2 Stir the suspension until the reaction is complete, about 18 hours. Remove the volatiles under vacuum, dissolve the residue in CH 3 OH and treat with excess Na 2

CO

3 plus NaHCO 3 Stir for 30 minutes and filter. Remove CH 3 OH from the filtrate, dissolve the residue in CH 3 CN, filter and remove CH 3 CN under reduced pressure. Dissolve the foam thus obtained in water, stir with an anion exchange resin such as IRA 401 S OH resin® (trademark of the Rohm and Haas Chemical Company, Philadelphia, Pennsylvania) for minutes. Filter and concentrate the aqueous solution, dissolve the residue thus obtained in CH 2

CI

2 and pass through a silica plug with 5% CH 3 OH in

CH

2

CI

2 as eluant. Concentrate the eluant and crystallize the resultant foam from CH 3 CN to obtain 0.9 g of the title compound, a solid.

1550; MS (FAB) 246 u I I III WO 94/19351 PCT/US94/01728 -42- Example 14 H 0 H

CH

3 N CH C HNAN N LCF3 H N N CF3 To a stirred solution of 86.5 g of the crude product of Preparation 5.1 in 1.3 L CH 3 CN under N 2 kept at -200C (water bath), add slowly 39 mL SOCI 2 Remove the water bath and stir vigorously at room temperature until complete reaction (about 12 to 18 hr) as judged by tic.

Concentrate under reduced pressure, take the resultant gum in a mixture of 1 L CH 2

CI

2 and 700 mL H 2 0, stir this mixture and neutralize to a pH of with slow addition of NH 4 0H. Filter this mixture to remove solids and wash the solids with CH 2

CI

2 Combine the filtrate and the washes, separate the layers, treat the aqueous layer with -1 g NaCI and then reextract with 2x500 mL CH 2

CI

2 Dry the combined organic layer (anhyd.

MgSO 4 and concentrate in vacuo to obtain 74 g of a light brown solid.

Slurry this solid in 225 mL EtOAc to obtain 35.6 g of off-white title compound. Concentrate the EtOAc filtrate and washes to obtain -37 g of a brown-black solid. Subject this solid to silica gel column chromatography to obtain an additional 3.6 g of product.

Chromatograph the above product on silica gel to obtain a light tan solid as an analytical sample of the title compound. M.p. 235- 237 0 C; MS 389 CI(NH 4 390 1040 (c MeOH).

Pharmaceutical Preparations The compounds of formula I can be combined with a suitable pharmaceutical carrier to prepare a pharmaceutical composition suitable for parenteral or oral administration. Such pharmaceutical compositions are useful in the treatment of cardiovascular and pulmonary disorders such as mammalian hypertension and bronchoconstriction.

The effective daily antihypertensive dose (ED50) of the present compounds will typically be in the range of about 1 to about 100 mg/kg of mammalian body weight, administered in single or divided doses. The exact dosage to be administered can be determined by the attending clinician and 3 7 1 I SWO 94/19351 PCT/US94/01728 43is dependent upon where the particular compound lies within the above cited range, as well as upon the age, weight and condition of the individual.

Generally, in treating humans in need of treatment for hypertension or bronchoconstriction, the present compounds can be administered in a dosage range of about 10 to about 500 mg per patient generally given a number of times per day, providing a total daily dosage of from about 10 to about 2000 mg per day.

The compositions of the present invention can be administered orally or parenterally. Typical injectable formulations include solutions and suspensions. Typical oral formulations include tablets, capsules, syrups, suspensions and elixirs. Also contemplated are mechanical delivery systems, e.g. transdermal dosage forms.

The typical acceptable pharmaceutical carriers for use in the formulations described above are exemplified by sugars such as lactose, sucrose, mannitol and sorbitol; starches such as cornstarch, tapioca starch and potato starch; cellulose and derivatives such as sodium carboxymethyl cellulose, ethyl cellulose and methyl cellulose; calcium phosphates such as dicalcium phosphate and tricalcium phosphate; sodium sulfate; calcium sulfate; polyvinylpyrrolidone, polyvinyl alcohol; stearic acid; alkaline earth metal stearates such as magnesium stearate and calcium stearate, stearic acid, vegetable oils such as peanut oil, cottonseed oil, sesame oil, olive oil and corn oil; non-ionic, cationic and anionic surfactants; ethylene glycol polymers; beta-cyclodextrin; fatty alcohols and hydrolyzed cereal solids; as well as other non-toxic compatible fillers, binders, disintegrants, buffers, preservatives, antioxidants, lubricants, flavo ig agents, and the like commonly used in pharmaceutical formulations.

Following are typical examples of oral and parenteral formulations, wherein the term "Active Ingredient" refers to a compound of formula I.

Capsule Amount (mg) Active Ingredient 250.0 125.0 Lactose 173.0 86.5 Corn Starch 75.0 37.5 Magnesium Stearate 2 TOTAL 500.0 250.0 Blend the active ingredient, lactose and corn starch until uniform; then blend the magnesium stearate into the resulting powder.

i I I- li~ WO 94/19351 PCT/US94/01728 -44- Encapsulate the mixture into suitably sized two-piece hard gelatin capsules.

Tablet Amount (mg) Active Ingredient 250.0 125.0 Lactose 161.0 80.5 Corn Starch 12.0 Water (per thousand tablets) 120 ml 60 ml (evaporates) (evaporates) Corn Starch 75.0 37.5 Magnesium Stearate 2.0 1.

TOTAL 500.0 250.0 Blend the active ingredient with the lactose until uniform.

Blend the smaller quantity of corn starch with the water and add the resulting corn starch paste then mix until a uniform wet mass is formed.

Add the remaining corn starch to the remaining wet mass and mix until uniform granules are obtained. Screen the granules through a suitable milling machine, using a 3/4 inch stainless steel screen. Dry the milled granules in a suitable drying oven until the desired moisture content is obtained. Mill the dried granules through a suitable milling machine using a 16 mesh stainless steel screen. Blend in the magnesium stearate and compress the resulting mixture into tablets of desired shape, thickness, hardness and disintegration.

Iniectable Solution ma/ml Active Ingredient 5.00 Methyl p-hydroxybenzoate 0.80 Propyl p-hydroxybenzoate 0.10 Disodium Edetate 0.10 Citric Acid Monohydrate 0.08 Dextrose 40.0 Water for injection qs. ad. 1.0 ml Dissolve the p-hydroxybenzoates in a portion of water for injection at a temperature of between 600C 70°C and cool the solution to 20°C 300C, Charge and dissolve all other excipients and the active ingredient. Bring the solution to final volume, filter it through a sterilizing membrane and fill into sterile containers.

I _1 .WO 94/19351 PCT/US94/01728 Biological Activity of 2-Benzyl Polycyclic Guanines The present compounds are useful in inhibiting the phosphodiesterase enzymes. These phosphodissterase enzymes are known to hydrolyze cGMP in smooth muscle. High levels of cGMP are associated with the relaxation of vascular smooth muscle, with a consequent subsequent reduction in blood pressure. Thus, it is believed that by inhibiting these phosphodiesterase enzymes, cGMP levels in muscle will be either maintained or increased, with a subsequent reduction in blood pressure. In vivo antihypertensive activity is determined orally in spontaneously hypertensive rats (SHR).

Phosphodiesterase inhibition in vitro: Compounds are evaluated for inhibition of two phosphodiesterase enzymes which hydrolyze cyclic guanosine monophosphate (cGMP). The first enzyme, calcium-calmodulin dependent phosphodiesterase (CaM-PDE), is a partially pure enzyme obtained from bovine aorta homogenates and purified by DEAE-cellulose and calmodulin-affinity chromatography. The enzyme is activated several fold by Ca-calmodulin and is selective for cGMP, although it will also hydrolyze cAMP. The second enzyme, cGMP phosphodiesterase (cGMP- PDE), is a homogeneous enzyme obtained from bovinE lung and purified by ion-exchange chromatography, gel filtration, and sucrose gradient centrifugation. cGMP-PDE is highly selective for cGMP. Bovine aorta homogenates and primary cultures of bovine aortic endothelial and vascular smooth muscle cells contain an enzyme with properties very similar to the lung isozyme.

The enzyme assay is performed using a Biomek Automated Pipetting Station. Compounds are dissolved in distilled water or DMSO and diluted with 10% DMSO. Compounds are tested at several concentrations at log intervals, typically 0.1, 1.0, 10, and 100 .M final concentration.

Assays contain the following components: 1 U.M substrate 3 H-cGMP mM Tris-HCI, pH 7.5, 5 mM MgCI2 0.5 mg/ml snake venom alkaline phosphatase 0.1 pM Calmodulin and 1 mM CaCl2 (for CaM-PDE only) Assays are initiated by addition of enzyme and stopped by addition of 10 mM isobutylmethylxanthine, a general phosphodiesterase inhibitor.

L LB~s~b dL 1 3 ~eLI WO 94/19351 PCT/US94/01728 -46- Assays are performed for 25 minutes at room temperature to achieve hydrolysis of substrate. The negatively charged substrates are then separated from guanosine by binding to an anion-exchange resin (AG1- X8) and centrifugation or filtration, and the product is quantitated by scintillation counting ir, counts per minute (cpm) of the remaining soluble material. Percent inhibition is calculated as follows: Inhibition= 100-[(cpm compound-blank)/(cpm control-blank)X100] Activity is expresssed as the ICso value, ie. the concentration required to inhibit activity of enzyme by 50 per cent.

Antihypertensive activity in rats The ability of the compounds of the present invention to lower blood pressure can be assessed in vivo in conscious spontaneously hypertensive rats (SHR). SHR males are purchased from Taconic Farms, Germantown New York and are approximately 16-18 weeks old when anesthetized with ether. The caudal (ventral tail) artery is cannulated with polyethylene tubing (PE50) and blood pressure and heart rate are recorded as described by Baum, T. et. al, J. Cardiovasc. Pharmacol. Vol pp. 655-667, (1983). Rats are placed into plastic cylindrical cages where they rapidly recover consciousness. Blood pressure and heart rate are allowed to stabilize for approximately 90 minutes prior to compound administration. Compounds are administered orally as solutions or suspensions in 0.4% aqueous methylcellulose vehicle via a feeding needle. The compound or 0.4% aqueous methylcellulose vehicle are given in a volume of 4 ml/kg to SHRs that had been fasted overnight.

Activity is expressed as the fall in mean blood pressure (MBP) in millimeters of mercury (mm Hg). Compound-induced changes are compared with the changes in an appropriate placebo group. "NT" means that the compound was not tested in that assay.

la _1 II~- WO 94/19351 WO 9419351PCTIUS94/01'728 -47- ACTIVITY OF 2-BENZYL-TETRACYLCI IC GUANINES f!E.Q~ SHR Athyeen Example CaM Dose Fall in MBP Number (U)(mpk) (mmHg) 1 0.1 10 47 1A 1 10 49 1I6 0.2 10 61 0.6 10 2 <0.1 10 2A 0.7 3 19 2B 0.4 3 3 1 10 21 3A 0.1 10 42 3 B 0.6 3 28 0.2 3 23 3 D NT 3 28 3 E 0.9 10 29 3F 0.8 10 34 3G 0.3 3 28 3 H 1 3 27 4 NT 3 32 NT 3 33 6 0.48 3 19 7 0.65 27 8 0.1 10 9 0.3 10 47 0.1 10 39 11 0.2 10 59 12 3.0 3 The activities of cis-5,6a,7,8,9,9a-hexahydro-5-i nethyl-2phenyl-3-(phenylmethyl)cyclopent[4, 5]imidazo[2, 1 blpurin-4(3H)-one (Cormpound A) and cis-5 ,6a,7, 8,9,9a-hexahydro-5-methylcyclopenta- 5]imidazo[2, 1-bI-purin-4(3H)-one (Compound compounds specifically disclosed in W091/1 971 7, are as follows: Cod. PDE-lC o: SR A CaM: 0.2 p.M fall in MBP at 25mpk: 6 mmHg B CaM: 33 p.M fafll in MBP at 25mpk: 32 mmHg fall in MBP at lompk: 8 mmHg

Claims (20)

1. A compound having the structural formula H H 3 CN I CH, R 2 Ra' R c R Rb or a pharmaceutically acceptable salt thereof, wherein: R 1 R 2 and R 3 are independently selected from the group consisting of hydrogen, lower alkyl, lower alkoxy, halogeno, hydroxy, (di- lower alkyl)amino, 4-morpholinyl, 1-pyrrolidinyl, 1-pyrrolyl, -CF 3 -OCF 3 phenyl and methoxyphenyl; or R 1 and R 2 together are methylenedioxy; or R 1 and R 2 together with the carbon atoms to which they are attached form a benzene ring; and R a is hydrogen and Rb and RC, together with the carbon atoms to which they are attached,.form a saturated ring of 5 carbons; or Ra is lower alkyl, Rb is hydrogen or lower alkyl, and Rc is hydrogen; or R a Rb and the carbon atom to which they are attached form a saturated ring of 7 carbons, ard Rc is hydrogen; or R a is hydrogen, and Rb, Rc and the carbon atoms to which they are attached form a tetrahydrofuran ring; or R a and Rb, together with the carbon atom to which they are attached, and Rb and Rc, together with the carbon atoms to which they are attached, each form a saturated ring of 5-7 carbons.

2. A compound of claim 1 wherein R 1 and R 3 are each hydrogen.

3. A compound of claim 1 wherein R 1 and R 3 are each hydrogen and R 2 is hydrogen, -CF 3 -OCF3, methyl, methoxy, fluoro, phenyl, methoxyphenyl, dimethylamino, 1-pyrrolidinyl or 1-pyrrolyl; or wherein R1 and R 2 together form a methylenedioxy group and R 3 is hydrogen.

4. A compound of any of claims 1, 2 or 3 wherein R a is hydrogen and Rb and Rc, together with the carbon atoms to which they are attached, form a saturated ring of 5 carbons; or R a is lower alkyl, Rb is ~p liar p IIR-Y~ 11~915P~IPI WO 94/19351 PCT/US94/01728 -49- hydrogen or lower alkyl, and RC is hydrogen; or R a Rb and the carbon atom to which they are attached form a saturated ring of 5 carbons, and RC is hydrogen; or R a is hydrogen, and Rb, RC and the carbon atoms to which they are attached form a tetrahydrofuran ring; or R a and Rb, together with the carbon atom to which they are attached, and Rb and Rc, together with the carbon atoms to which they are attached, each form a saturated ring of carbons. A compound of any of claims 1, 2 or 3 wherein R a is lower alkyl, Rb is lower alkyl or hydrogen, and Rc is hydrogen; R a and Rb and the carbon atom to which they are attached form a saturated ring of carbons and Rc is hydrogen; or R a is hydrogen and Rb and Rc, together with the carbon atoms to which they are attached, form a saturated ring of carbons.

6. A compound of claim 1 having the structural formula wherein R a is hydrogen and Rb and Rc, together with the carbon atoms to which they are attached, form a saturated ring of 5 carbons, and wherein R 1 R 2 and R 3 are as defined in the following table: R1 R 2 Rs H H H -OCH 3 H H H F H H -OCH 3 H H OH H H -CH 3 H H (CH 3 2 N- H -OCH 3 -OCH 3 -OCH 3 -OCH3 -OCH3 H -CF 3 H H H CeH5- H -1 9 11 R2 R3 H -OCFa H H NC H 3,4-OCH2O H RI _R2 R3 H CoH H -JO.CHj H III and R2, together with the H carbon atoms to which they are attached form a benzene ringi H Cl a. a a. C a C C C C C a C. C C.. 06 C C 0 C C C 0. C.

7. A compound of claim 1 selected from the group consisting of: 5'-methyl-2'-(phenylmethyl)-spiro[cyclopentane-1 imidazo[2, 1 purinl-4'(5'H) -one; 5,'7,7-trimethyl-2-(phenymethyl)-3H-imidazol[2, 1 one; )-2-benzyl-7, 8-dihydro-5-methyl-7-( 1 -methylethyl)- 1 H- imidazo[2, 1 7, 8, 9, 9a, 10, 11, 11la-octahydro-5-methyl-2-(3,4- methylene-dioxyphenylmethyl)-3H-pentalen[6a, 1:4, Slimidazo[2, 1-blpurin- 4(5H)-one; and )-cis-6a, 7, 9 9a-tetrahydro-5-methyl-2- [4-(trifluoromethyl)- phenylmethyl]-3H-furo[3', 4':4,5]imidazo[2, 1-blpurin-4(5H)-one.

8. A pharmaceutical composition comprising an effective amount of a compound as defined in any one of claims 1 to 5 in a pharmaceutically acceptable carrier. S:',I6104GW 7 ~s i 51

9. A pharmaceutical composition comprising an effective amount of a compound of any of claims 6 or 7 in a pharmaceutically acceptable carrier. A process for the preparation of a pharmaceutical composition as claimed in claim 8 which comprises admixing a compound as defined in any one of claims 1 to 5 with a pharmaceutically acceptable carrier.

11. A method of treating hypertension, angina, bronchoconstriction, restenosis post angioplasty, atherosclerosis, ischemia, peripheral vascular diseases, or diseases benefitting from platelet inhibition, or for maintaining guanosine monophosphate (cGMP) levels, comprising administering an effective amount of a compound of claim 1 to a mammal in need of such treatment.

12. A process for preparing a polycyclic guanine of formula II: 0 R 4 H N N R(CH2)n Rb 1 RoM Rl II wherein: SR4 is H, aikyl or alkyl substituted with aryl or -OH; R 5 is H. halo, -CF 3 alkoxy, alkylthio, alkyl, cycloalkyl, -SOgNH 2 -NH 2 monoalkylamino, dialkylamino, hydroxyalkylamino, aminoalkyl- amino, -COOH, alkoxycarbonyl, aminocarbonyl, arly, substituted aryl, or alkyl substituted with aryl, substituted aryl, -OH, alkoxy, -NH 2 monoalkylamino or dialkylamino; Ral, Rbl, R cl and Rdl independently represent H, alkyl, cycloalkyl or aryl; or (Ral and Rbl) or (Rol and Rdl) or (Rbl and Rol) can complete a saturated ring of 5- to 7- carbon atoms, or (Ral and Rbl) taken together and (Rbl and R 1 o) taken together, each complete a saturated ring of 5- to 7-carbon atoms, wherein each ring optionally can contain a sulfur or oxygen atom and whose carbon atoms may be optionally substituted with one or more or the following: alkenyl, alkynyl, -OH, -COOH, S:16104GW C I. C- C- L L_ I I WO 94/9351 PCT/US94/01728 -52- alkoxycarbonyl, alkyl or alkyl substituted with -OH, -COOH or alkoxycarbonyl; or such saturated ring can have two adjacent carbon atoms which are shared with an adjoining aryl ring; and n is zero or one; comprising a) reducing a nitrosopyrimidine of formula I: 0 R4 1 NO III 016 Z I NH 2 wherein R 4 is as defined above, Z is =0 or -OR 6 wherein R 6 is alkyl; and the dotted lines indicate an optional double bond such that when Z is position 1 contains hydrogen and there is single bond between positions 1 and 2 on the ring, or when Z is -OR 6 wherein R 6 is alkyl, there is a double bond between positions 1 and 2 on the ring, and treating the reduced nitrosopyrimidine with in acylating reagent of the formula R 5 COL (IV) wherein R 5 is as defined above, and L represents a leaving group, to give the amidopyrimidine V: O S 4 5 NHCOR s 6 V Z N NH2 wherein R 4 R 5 the dotted lines and Z are as defined above; b) reacting amidopyrimidine V with an effective amount of a halogenating/cyclizing reagent to give halopurine of formula VI: O R 4 H R"t V I wherein R 4 and R 5 are as defined above, and X is halo; c) reacting, in the presence of a base, the halopurine with an amine of the formula VII: H 2 N (CH 2 )n -OH Ralb C Rbi RI R VI '~sNNW. IY II 1~~ -53- wherein Ral, Rbl, Re l Rdl and n are as defined above to give th substituted aminopurine VIII: O R 4 l H HN N (CH2)n-OH 1hRC- d VIII wherein R 4 R 5 Rbl, R b R,1 Rdl and n are as defined above; and d) closing the ring of substituted aminopurine with a suitable dehydrating agent.

13. The process of claim 12, wherein in step the reduced nitrosopyrimidine is treated with an acylating reagent in the presence of either an acylating catalyst and/or coupling reagent and/or phase transfer catalyst.

14. A process for preparing an amidopyrimidine of formula V: O 4L NHCORs Z NH2 wherein R 4 R 5 the dotted lines and Z are as defined in claim 12, comprising: t*f reducing a nitrosopyrimidine (iI) as defined in claim 12, *i and treating the reduced nitrosopyrimidine with an acylating reagent. at a The process of claim 14, wherein the reduced nitroso- pyrimidine is treated with acylating reagent in the presence of either an acylating catalyst and/or coupling reagent and/or phase transfer catalyst.

16. A process for preparing a halopurine of formula VI: 0 O H iR VI wherein, wherein, r~ III I--ia I, l'yl= i I III -54- R 4 is hydrogen, alkyl or alkyl substituted with aryl or hydroxy; R 5 is hydrogen, halo, trifluoromethyl, alkoxy, alkyithlo, alkyl, cycloalkyl, aminosulfonyl, amino, monoalkylamino, dialkylamino, hydroxyalkylamino, aminoalkylamino, carboxy, alkoxycarbonyl, amino- carbonyl, aryl, substituted aryl, or alkyl substituted with aryl, substituted aryl, hydroxy, alkoxy, amino, monoalkylamino or dialkylamino; and X is halo; comprising reacting an amidopyrimidine as defined in claim 12 With an effective amount of a halogenating/cyclizing reagent.

17. The p;ocess of claim 16 wnerein the amidopyrimidine is reacted with an effective amount of a halogenating/cyclizing reagent in the presence of either one or more additional halide sources and/or a phase transfer catalyst to give the haiopurine (VI).

18. The process of claim 16.further comprising the step of: reacting, in the presence of a base, a halopurine (VI) with an amine of the formula VII: HgN (CH2)n-OH Rai Rb1 o Rdl VII wherein Ral, Rbl, Rd, Rdl and n are as defined in claim 12 to give the substituted aminopurine (VII) as defined in claim 12,. i 19. The process of claim 18 further comprising the step of: closing the ring of substituted aminopurine VIII with a suitable dehydrating agent to give a polycyclic guanine (II) as defined in claim 12. A pa .cess for preparing a substituted aminoputine of formula VIII: R4 H N HN Ri (CH 2 )n-OH aa Rb1Rcl Rd' VIII wherein R 4 is hydrogen, alkyl or alkyl substituted with aryl or hydroxy; I' I I i r or R 5 is hydrogen, halo, trifluoromethyl, alkoxy, alkylthio, alkyl, cycloalkyl, aminosulfonyl, amino, monoalkylamino, dialkylamino, hydroxyalkylamino, aminoalkylamino, carboxy, alkoxycarbonyl, aminocarbonyl, aryl, substituted aryl, or alkyl substituted with aryl, substituted aryl, hydroxy, alkoxy, amino, monoalkylamino or dialkylamino; R al Rbl, R l and Rdl Independently represent hydrogen, alkyl, cycloalkyl or aryl; or (Ral and Rbl) or (R 01 and Rdl) or (Rbl and R 01 can complete a saturated ring of 5- to 7- carbon atoms, or (Ral and Rbl) taken together and (Rbl and R cl taken together, each complete a saturated ring of 5- to 7-carbon atoms, wherein each ring optionally can contain a sulfur or oxygen atom and whose carbon atoms may be optionally substituted with one or more or the following: alkenyl, alkynyl, hydroxy, carboxy, alkoxycarbonyl, alkyl or alkyl substituted with hydroxy, carboxy or alkoxycarbonyl; or such saturated ring can have two adjacent carbon atoms which are shared with an adjoining aryl ring; and n is zero or one, comprising reacting halopurine (VI) as defined in claim 12 with an amine of the formula VII: \1 H (CH2)n OH Rd Rl vd wherein Ral, Rbl, R cl Rdl and n are as defined above.

21. The process of claim 20 ,wherein halopurine VI is reacted with amine VII in the presence of an added base, to give substituted aminopurine VIII.

22. The process of claim 20 further comprising the step of: closing the ring of substituted aminopurine VIII with a suitable dehydrating agent to give polycyclic guanine II.

23. Amidopyrimidine V: .o. O NHCOR ZNH 2 wherein R 4 is hydrogen, alkyl or alkyl substituted with aryl or hydroxy; II -56- R 5 is alkyl substituted with aryl, ;ii substituted aryl, hydroxy, alkoxy, amino, monoalkylamino or dialkylamino; Z is =0 or -OR 6 wherein R 6 is alkyl; and the dotted lines indicate an optional double bond such that ;I when Z is position 1 contains hydrogen and there is single bond i between positions 1 and 2 on the ring, or when Z is -OR 6 wherein R 6 is alkyl, there is a double bond between positions 1 and 2 on the ring, with the proviso that when Z is R 4 and R s cannot both be methyl.

24. A process for preparing a compound of formula I as defined in claim 1 comprising a) cyclizing an aminopurine of the formula OH o o ,o compound of formula I; or b) reducing a polycyclio guanine of the formulall L x. o wh re A3 S" R OH-- OH i wherein R1, R2, R3, R a Rb and RC are as defined in claim 1, to obtain a compound of formula I; or c) reacting a tetracyclic guanine of the formula 9i 'A if~ 4JJ PC- -r 57 S>-Br with a compound of the formula Ar-CH 2 -ZnBr, wherein Ar is optionally substituted phenyl or naphthyl, to obtain a compound of formula I wherein Ra is hydrogen and Rb and Rc, together with the carbon atoms to which they are attached, form a saturated ring of 5 carbons. A compound as defined in claim 1 or a pharmaceutically acceptable salt thereof, and substantially as described herein with reference to the Examples.

26. A process for preparing a polycyclic guanine of formula II as defined in claim 12, and substantially as described herein with reference to the Examples. :".DATED this 16th day of June 1997 SCHERING CORPORATION By their Patent Attorneys GRIFFITH HACK 9 0* 1 C' S:16104GW t i INTERNATIONAL SEARCH REPORT Applic Intmr',mal Applicaon No PCi/US 94/01728 A. CLASSIFICATION OF SUBJECT MA'IT.ER IPC 5 C07D487/14 A61K31/505 C07 487/20 C07D491/22 0070473/18 C07D473/40 C070239/54 0070239/52 //(C07D487/14,239:00, 235:00,235:00),(C07D487/20,239:00,235:00,235:00),(C07D491/22, According to Intcmatonal Patent Classification (IPC) or to hth national classification and IPC B. FIELDS SEARCHED Minimum documentation searched (classification system followed by classification symbols) IPC 5 C07D A61K Documentation se.hed other than minimum documentation to the extent that such documents are included in the fields searched Electronic data base consulted dunng the intcmatonal search (name of data base and, where practcal, search terms used) C. DOCUMENTS CONSII)ERED TO BE RELEVANT Category Citation of document, with indication, where appropriate, of the relevant passages Relevant to claim No. X WO,A,91 19717 (SCHERING) 26 December 1991 1,9,10 cited in the application see claims 1,9 X EP,A,O 466 164 (BOEHRINGER) 15 January 1992 see claim 1 X US,A,4 663 446 WRIGHT) 5 May 1987 see column 5, line 13 SFurther documents arc listed in the continuaton of box C. Patent family members are listed n annex. Spenal categories of cited documents Special categories of cited documents: T later document published after the international filing date A doc defin the or priority date and not in conflict with the application but document defining the general state of the art which is notto understand the principle or theory uniidelying the considered to he of particular relevance invention earlier document but published on or after the international docmnent of particular relevance; the claimed invention filing date cannot be considered novel or cannot be considered to document which may throw doubts or pronty claim(s) or involve an inventve step when the document is taken alone which is cited to establish the publication date of another document of partcular relevance; the claimed nvention tation or other special reason (as specified) cannot be considered to involve an inventive step when the "0 document referring to an oral disclosure, use, exhibition or document is combined with one or more other such docu- other means ments, such combination being obvious to a person skilled document published pnor to the incrnational filing date but in the art. later than the pnority date claimed document member of the same patent family Date of the actual completion of the international search Date of mailing of the internauonal search report 14 June 1994 29. 06.9 Name and mailing address of the ISA Authonzed officer European Patent Office, P.B. 5818 Patenllan 2 NL 2280 IIV Rilswilk Tel.(+31-70)340.2040,Tx.31651 poni, Alfaro Faus I a 3170) 340-3016 A aro aus, Form PCT/ISA/210 (seond sheet) (July 1992) page 1 of INTERNATIONAL SEARCH REPORT' nter,-minal ApplicttUn NO PC /US 94/01728 f A. CLASSIFICATION OF SUBJ.EC' MA'rrER IPC 5 307:00,239:00,235:00,235:00) According to Internatonal Patent Classification (11C) or to both natonal classification and IPC B. FIELDS SEARCHED Minimum documentation searched (classification system followed by classification symbols) Documentation searched other than minimum documentation to the extent that such documents are included in the fields searched Electronic data base consulted dunng the international search (name of data base and, where practical, search terms used) C. DOCUMENTS CONSIDERED TO li R.EEVANT______ Category' Citation of document, with indication, where appropriate, of the relevant passages Relevant to claim No. X CHEMICAL ABSTRACTS, vol. 81, no. 11, 1974, Columbus, Ohio, US; abstract no. 62626q, H. BEERBUUM ET AL. 'Mass spectrometric studies of halogenated purines' page 391 see compound II Z. CHEM. 1974, 14(4), 156 7 r Further documents are listed in the continuation of box C. j Patent family members arc listed in annex. Special categories of cated documciis: Sp;al categores of cited docu s T later document published after the internatonal filing date or pnonty date and not in conflict with Lhe application but document defining the general state of the art which is not a ed to understand the pnncple or theoy underlying the considered to be of particular relevance invention E earlier document but published on or after the international "X document of particular relevance; the claimed invention filing dite cannot be considered novel or cannot be considered to document which may throw doubts on pnority claim(s) or involve an inventive step when the document is taken alone whih is cited to establish the publication date of another document of particular relevance; the claimed invention citation or other special reason (as specified) cannot be considered to involve an inventive step when the document referring to an oral disclosure, use, exhibition or document is combined with one or more other such docu- other means ments, such combination being obvious to a person skilled document published prior to the international filing date but in the art. later than the pnonty date claimed document member of the same patent faminuly Date of the actual completion of the international search Date of mailing of the internatonal search report 14 June 1994 Name and mailing address of the ISA Authonzed officer European Patent Office, P.D. 5818 Patentlaan 2 NL 2280 HV RijswiAk Tel. (+31-70) 340-040, Tx. 31 6S1 epo Alfaro Faus, I Fax: 1 31-70) 340.16 Alfaro aus, Form PCT/ISA/210 (ie.owd sheet) (July 1991) page 2 of INTERNATIONAL SEARCH RPORIT Fntriiona Apaion nNNo PCi/US 94/01728 C(Continuation) DOCUMENTS CONSIDERED TO BiE RELEVANT Category' Citation of document, with indication, where appropriate, of the relevant passages Relevant to claim No. X CHEMICAL ABSTRACTS, vol. 98, no. 21, 26 1983, Columbus, Ohio, US; abstract no. 174511f, 'Formation of cyclic 1,N2-adducts by reaction of deoxyguanosine with alpha-acetoxy-N-nitrosopyrrolidine, 4-(carbethoxynitrosoamino)butanal, or crotonaldehyde' page 238; see [85352-98-3! CANCER RES. 1983, 43(3), 1230 X CHEMICAL ABSTRACTS, vol. 116, no. 21, 26 1992, Columbus, Ohio, US; abstract no. 209475s, M. DYPBUKT ET AL. 'Cytotoxic and genotoxic effects of styrene-7,8-oxide in neuroadrenergic Pc 12 cells' page 252; see abstract and chemical substance index, page 8641, column 1, lines 118 121 CARCINOGENESIS (LONDON) 1992, 1L3(3), 417 424 X CHEMICAL ABSTRACTS, vol. 110, no. 1, 26 1989, Columbus, Ohio, US; abstract no. 2572b, K. HEMMINKI ET AL. 'Styrene oxide as a stereochemical probe for the mechanism of aralkylation at differert sites on guanosi nel page 240; see abstract and 12th chemical substance collective index, page 77869, column 2, lines 38 41 CHEM. RES. TOXICOL. 1988, 1(6),364-9 X EP,A,0 039 780 (NESTLE) 18 November 1981 24 see page 9, line 16 line X LIEBIGS ANNALEN DER CHEMIE 1974 WEINHEIM 24 DE pages 2030 2045 W. PFLEIOERER 'Synthese, Struktur und Eigenschaften der Harnsdure sowie ihrer N-Methylderivare' see page 2032, compound 1 Form PCTIISAJ21 (continuation of Second8 sheet) (July 1992) page 3 of 1Tl'rr.D ATTAXIAl QVA001.1 IM13fAUT 6 "kh- 1kIt7 K!TIThA (~7ADCI t~)Cl) I N L1~N( I I iA~..A AA~ ~Intem-mal Appieattoii No PCiIUS 94/01728 C(Continuauon) DOCUMENTS CONSIDEREDT) 1 REiULEVANT Category' Citation of document, with indication, where appropnate, of the relevant paiisagcs Relevant to claim No. X CHEMICAL ABSTRACTS, vol. 99, no. 1, 24 1983, Columbus, Ohio, US; abstract no. 100b, F. GASPARI ET AL. 'Apparent dissociation constants of some possible uracil metabolites of methylxanthines' page 9 see abstract and 11th Chemical Substance Collective Index, page 106, column 1, lines 94-96, 110 and page 28108, column 3, lines 44 46 and 54 ANAL. LETT. 1983, 16(B3), 167 X JOURNAL OF THE CHEMICAL SOCIETY 1964 ,24 LETCHWORTH GB pages 565 572 F. BERGMANN ET AL. 'Formation of 6-hydroxypteridines by condensation of with chloral' see page 571, paragraph 3 X EP,A,O 184 738 THOMAE) 18 June 1986 24 see examples 5a, 19a, 21a, 22a and 23a X EP,A,O 149 200 THOMAE) 24 July 1985 24 see example 28a X CHEMICAL ABSTRACTS, vol. 67, no. 1967, 24 11 September 1967, Columbus, Ohio, US; abstract no. 54153d, M. SEKIYA ET AL. 'Derivatives of uracil or xanthine' page 5098 see abstract JP,A,8 621 (YOSHITOMI) 18 April 1967 X CHEMICAL ABSTRACTS, vol. 108, no. 11, 24 1988, Columbus, Ohio, US; abstract no. 94267r, K. MAMADA ET AL. 'Synthesis of deuterium-labeled 1,3-dimethyluric acids' page 631; see abstract J. LABELLED COMP. RADIOPHARM. 1987, 24(4) ,361-8 X JOURNAL OF HETEROCYCLIC CHEMISTRY 24 vol. 3 1966 PROVO US pages 476 481 J. FU ET AL. '8-Arylpuirines and their ultraviolet absorption spectra' see page 477, compound V Form PCTIISA.'l 0 (continuation of gecand Ilicet) (July 1992) page 4 of lN'rr-RNA'I'IONA1. SLARk1 RtK1UN I ntematiional Application No jPC,, JS 94/01728 C.(Continuation) DOCUMMNS CONSIDERED TO li REJLEVANTr Category' Ci tation of documentl, with indication, where appropniate -or the relevant pamags Relevant to claim No. P,X JOURNAL OF MEDICINAL CHEMISTRY 24 vol. 36, no. 10 1993 WASHINGTON US pages 1465 1473 L. FRAISE ET AL. 'Long-chain-substituted uric acid and 5,6-diaminouracil derivatives as novel agents agains free radical processes: synthesis and in vitro activity' see page 1471, compounds 3a 3d P,X JOURNAL OF MEDICINAL CHEMISTRY 24 vol. 36, no. 22 1993 WASHINGTON US pages 3341 3349 C. E. MULLER ET AL. 'Synthesis of paraxanthine analogs (1,7-disubstituted xanthines) and other xanthines unsubstituted at the 3-position: structure-activity relationships at adenosine receptors' see -age 3348, compounds 29b, P,X CHEMICAL ABST, 'TS, vol. 119, no. 19, 26 1993, Columbus, Ohio, US; abstract no. 197329u, J.R. CUSHNIR El AL. 'Tasndem mass spectrometric approaches foe the analysis of alkylguanines in human urine' page~ 270; see abstract ORG. MASS SPECTROM. 1993, 28(5), 552-8 Form PCTJISA210 (continuation of scond sheet) (July 1992) page 5 of INTERNATIO'AL SEIARtCH RlIOR10'I lntm-rinAiApplication No .fornation on patent family members PC i,tJS 94/01728 Patent document Publication IPatent family I Publication cited in search report date member(s) date WO-A-9119717 26-12-91 AU-A- 8199291 07-01-92 EP-A- 0538332 28-04-93 NZ-A- 238609 23-12-93 EP-A-0466164 15-01-92 DE-A- 4022314 16-01-92 CA-A- 2046846 14-01-92 JP-A- 4230383 19-08-92 US-A- 5264573 23-11-93 US-A-4663446 05-05-87 NONE EP-A-0039780 18-11-81 CH-A- 643260 30-05-84 AU-B- 539258 20-09-84 AU-A- 6961781 05-11-81 CA-A- 1155841 25-10-83 GB"-A,B 2075505 18-11-81 56169690 26-12-81 US-A- 4469698 04-09-84 US-A- 4581451 08-04-86 EP-A-0184738 18-06-86 OE-A- 3445299 19-06-86 JP-A- 61145183 02-07-86 EP-A-0149200 24-07-85 DE-A- 3347290 11-07-85 AU-A- 3721184 04-07-85 JP-A- 60172980 06-09-85 US-A- 4722929 02-02-88 JP-A-862 1 NONE Form PCT/ISA&i210 (patent family annex) (July 1992)